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Journal articles on the topic "Pab1"
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Ardelean, Radu, Adriana Popa, Ecaterina Stela Drăgan, Corneliu-Mircea Davidescu, and Maria Ignat. "New Polymeric Adsorbents Functionalized with Aminobenzoic Groups for the Removal of Residual Antibiotics." Molecules 27, no. 9 (2022): 2894. http://dx.doi.org/10.3390/molecules27092894.
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In this paper, we present the synthesis of new polymeric adsorbents derived from macroporous chloromethylated styrene–divinylbenzene (DVB) copolymers with different cross-linking degrees functionalized with the following aminobenzoic groups: styrene—6.7% DVB (PAB1), styrene—10% DVB (PAB2), and styrene—15% DVB (PAB3). The new polymeric products, PAB1, PAB2, and PAB3, were characterized by FTIR spectroscopy, thermogravimetric analysis, and EDX, SEM, and BET analysis, respectively. The evolution of the functionalization reaction was followed by FTIR spectroscopy, which revealed a decrease in the intensity of the γCH2Cl band at 1260 cm−1, and, simultaneously, the appearance of C=O carboxylic bands from 1685–1695 cm−1 and at 1748 cm−1. The thermal stability increased with the increase in the cross-linking degree. The data obtained from the EDX analysis of the novel cross-linked copolymers confirmed the functionalization with aminobenzoic groups through the presence and content of nitrogen, as follows: PAB1: N% = 0.47; PAB2: N% = 0.85; and PAB3: N% = 1.30. The adsorption performances of the novel polymeric adsorbents, PAB1, PAB2, and PAB3, were tested in the adsorption of three antibiotics, tetracycline, sulfamethoxazole, and amoxicillin, from aqueous solutions, by using extensive kinetic, equilibrium, and thermodynamic studies. The best adsorption capacity was demonstrated by the tetracycline. Amoxicillin adsorption was also attempted, but it did not show positive results.
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Amirkhosravi, Ali, Todd V. Meyer, Liza Robles-Carrillo та ін. "β2-Glycoprotein 1 Antibodies Directly Activate the Platelet IgG Receptor, FcγRIIa, and Cause Thrombosis in FCGR2A Transgenic but Not in Wild Type Mice". Blood 120, № 21 (2012): 106. http://dx.doi.org/10.1182/blood.v120.21.106.106.
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Abstract Abstract 106 Antibodies targeting β2-glycoprotein 1 (β2-GP1; β2-Abs) are of primary importance in antiphospholipid syndrome (APS), a thrombotic autoimmune disorder. The predominance of the IgG antibody isotype in APS is conspicuously associated with increased risk of thrombosis, raising the question whether the platelet IgG receptor, FcγRIIa, may play a role in thrombosis in APS, as is the case in heparin-induced thrombocytopenia (HIT). The hypothesis that platelet FcγRIIa may contribute to thrombosis in APS has received little attention, with research emphasis instead placed on several proposed alternative mechanisms of action. We have shown that, like HIT antibodies, antibodies targeting VEGF or CD154 are also potently thrombotic in mice transgenic for human FcγRIIa but have no activity in mice lacking FcγRIIa (i.e., wild type mice). We therefore asked whether antiphospholipid antibodies can activate platelets and cause thrombosis via FcγRIIa. To this end, we tested mouse monoclonal (mAb) and goat polyclonal anti-human β2-Abs alone or complexed to human β2-glycoprotein 1 (i.e., to form immune complexes, or ICs) by the serotonin release assay (SRA) and platelet aggregation methods using washed human platelets. We found that two of three commercially available polyclonal anti-β2-Abs (pAb1 and pAb3), both alone or in IC form, induced platelet granule release and aggregation, and that this activity was abolished by anti-FcγRIIa mAb, IV.3. pAb2 and mAb were inactive. Activity analysis (SRA) of preformed ICs using constant pAb1 concentration with varied β2-GP1 stoichiometries revealed a zone-of-equivalence pattern, with maximal activity near balanced stoichiometry (1:1). Because pAb2 did not activate platelets, we sought to determine its capacity to form higher order ICs (which are known to be required for FcγRIIa activation) by size exclusion chromatography (SEC). All antibodies tested in isolation were shown by HPLC-SEC to be free of aggregates or degradation products. Antibody-antigen complex size analysis revealed that, as expected, mAb+β2-GP1 in 1:1 stoichiometry failed to form higher order ICs (i.e., complexes having ≥2 IgGs/complex). pAb2 (inactive) did form higher order ICs at 1:1 and 4:1 (IgG:Ag) stoichiometries, but less extensively than pAb1 (active), which efficiently formed higher order ICs at both 1:1 and 4:1 stoichiometries. pAb1 alone (i.e., not in IC form) caused aggregation, while pAb2 did not. Furthermore, preincubation of washed platelets with pAb2 prevented pAb1-induced aggregation, suggesting that pAb1 activity is β2-GP1-specific. A single intravenous injection of anti-β2-GP1 ICs (20 μg β2-GP1 plus 120 μg pAb1, a 1:2 molar stoichiometry) induced severe thrombocytopenia (>90%) and caused thrombotic shock in FcγRIIa-transgenic mice but not in wild type mice. Symptoms of shock occurred within 10 minutes. Pervasive occlusive thrombi were observed in the lungs of all FcγRIIa-transgenic but not in any wild type mice (H&E microscopy). Injection of pAb2 ICs produced none of these effects in transgenic or wild type mice. Finally, injection of pAb1 or pAb2 alone (neither of which bind mouse β2-GP1) had no effects in transgenic mice. In summary, these findings confirm previous in vitro studies that β2-Abs can directly activate platelets in a manner wholly dependent on the platelet IgG receptor, FcγRIIa. Additionally, we have shown for the first time in vivo that β2-Abs can also cause FcγRIIa-dependent thrombosis. This mechanism may contribute to thrombosis in APS, suggesting that further studies on the importance of FcγRIIa in APS are warranted. Disclosures: No relevant conflicts of interest to declare.
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Mangus, David A., Nadia Amrani, and Allan Jacobson. "Pbp1p, a Factor Interacting withSaccharomyces cerevisiae Poly(A)-Binding Protein, Regulates Polyadenylation." Molecular and Cellular Biology 18, no. 12 (1998): 7383–96. http://dx.doi.org/10.1128/mcb.18.12.7383.
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ABSTRACT The poly(A) tail of an mRNA is believed to influence the initiation of translation, and the rate at which the poly(A) tail is removed is thought to determine how fast an mRNA is degraded. One key factor associated with this 3′-end structure is the poly(A)-binding protein (Pab1p) encoded by the PAB1 gene inSaccharomyces cerevisiae. In an effort to learn more about the functional role of this protein, we used a two-hybrid screen to determine the factor(s) with which it interacts. We identified five genes encoding factors that specifically interact with the carboxy terminus of Pab1p. Of a total of 44 specific clones identified,PBP1 (for Pab1p-binding protein) was isolated 38 times. Of the putative interacting genes examined, PBP1 promoted the highest level of resistance to 3-aminotriazole (>100 mM) in constructs in which HIS3 was used as a reporter. We determined that a fraction of Pbp1p cosediments with polysomes in sucrose gradients and that its distribution is very similar to that of Pab1p. Disruption ofPBP1 showed that it is not essential for viability but can suppress the lethality associated with a PAB1 deletion. The suppression of pab1Δ by pbp1Δ appears to be different from that mediated by other pab1 suppressors, since disruption of PBP1 does not alter translation rates, affect accumulation of ribosomal subunits, change mRNA poly(A) tail lengths, or result in a defect in mRNA decay. Rather, Pbp1p appears to function in the nucleus to promote proper polyadenylation. In the absence of Pbp1p, 3′ termini of pre-mRNAs are properly cleaved but lack full-length poly(A) tails. These effects suggest that Pbp1p may act to repress the ability of Pab1p to negatively regulate polyadenylation.
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Dufresne, Philippe J., Eliane Ubalijoro, Marc G. Fortin, and Jean-François Laliberté. "Arabidopsis thaliana class II poly(A)-binding proteins are required for efficient multiplication of turnip mosaic virus." Journal of General Virology 89, no. 9 (2008): 2339–48. http://dx.doi.org/10.1099/vir.0.2008/002139-0.
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The poly(A)-binding protein (PABP) is an important translation initiation factor that binds to the polyadenylated 3′ end of mRNA. We have previously shown that PABP2 interacts with the RNA-dependent RNA polymerase (RdRp) and VPg-Pro of turnip mosaic virus (TuMV) within virus-induced vesicles. At least eight PABP isoforms are produced in Arabidopsis thaliana, three of which (PABP2, PABP4 and PABP8) are highly and broadly expressed and probably constitute the bulk of PABP required for cellular functions. Upon TuMV infection, an increase in protein and mRNA expression from PAB2, PAB4 and PAB8 genes was recorded. In vitro binding assays revealed that RdRp and the viral genome-linked protein (VPg-Pro) interact preferentially with PABP2 but are also capable of interaction with one or both of the other class II PABPs (i.e. PABP4 and PABP8). To assess whether PABP is required for potyvirus replication, A. thaliana single and double pab knockouts were isolated and inoculated with TuMV. All lines showed susceptibility to TuMV. However, when precise monitoring of viral RNA accumulation was performed, it was found to be reduced by 2.2- and 3.5-fold in pab2 pab4 and pab2 pab8 mutants, respectively, when compared with wild-type plants. PABP levels were most significantly reduced in the membrane-associated fraction in both of these mutants. TuMV mRNA levels thus correlated with cellular PABP concentrations in these A. thaliana knockout lines. These data provide further support for a role of PABP in potyvirus replication.
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Yao, Gang, Yueh-Chin Chiang, Chongxu Zhang, Darren J. Lee, Thomas M. Laue, and Clyde L. Denis. "PAB1 Self-Association Precludes Its Binding to Poly(A), Thereby Accelerating CCR4 Deadenylation In Vivo." Molecular and Cellular Biology 27, no. 17 (2007): 6243–53. http://dx.doi.org/10.1128/mcb.00734-07.
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ABSTRACT The mRNA deadenylation process, catalyzed by the CCR4 deadenylase, is known to be the major factor controlling mRNA decay rates in Saccharomyces cerevisiae. We have identified the proline-rich region and RRM1 domains of poly(A) binding protein (PAB1) as necessary for CCR4 deadenylation. Deletion of either of these regions but not other regions of PAB1 significantly reduced PAB1-PAB1 protein interactions, suggesting that PAB1 oligomerization is a required step for deadenylation. Moreover, defects in these two regions inhibited the formation of a novel, circular monomeric PAB1 species that forms in the absence of poly(A). Removal of the PAB1 RRM3 domain, which promoted PAB1 oligomerization and circularization, correspondingly accelerated CCR4 deadenylation. Circular PAB1 was unable to bind poly(A), and PAB1 multimers were severely deficient or unable to bind poly(A), implicating the PAB1 RNA binding surface as critical in making contacts that allow PAB1 self-association. These results support the model that the control of CCR4 deadenylation in vivo occurs in part through the removal of PAB1 from the poly(A) tail following its self-association into multimers and/or a circular species. Known alterations in the P domains of different PAB proteins and factors and conditions that affect PAB1 self-association would, therefore, be expected to be critical to controlling mRNA turnover in the cell.
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Amrani, N., M. Minet, M. Le Gouar, F. Lacroute, and F. Wyers. "Yeast Pab1 interacts with Rna15 and participates in the control of the poly(A) tail length in vitro." Molecular and Cellular Biology 17, no. 7 (1997): 3694–701. http://dx.doi.org/10.1128/mcb.17.7.3694.
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In Saccharomyces cerevisiae, the single poly(A) binding protein, Pab1, is the major ribonucleoprotein associated with the poly(A) tails of mRNAs in both the nucleus and the cytoplasm. We found that Pab1 interacts with Rna15 in two-hybrid assays and in coimmunoprecipitation experiments. Overexpression of PAB1 partially but specifically suppressed the rna15-2 mutation in vivo. RNA15 codes for a component of the cleavage and polyadenylation factor CF I, one of the four factors needed for pre-mRNA 3'-end processing. We show that Pab1 and CF I copurify in anion-exchange chromatography. These data suggest that Pab1 is physically associated with CF I. Extracts from a thermosensitive pab1 mutant and from a wild-type strain immunoneutralized for Pab1 showed normal cleavage activity but a large increase in poly(A) tail length. A normal tail length was restored by adding recombinant Pab1 to the mutant extract. The longer poly(A) tails were not due to an inhibition of exonuclease activities. Pab1 has previously been implicated in the regulation of translation initiation and in cytoplasmic mRNA stability. Our data indicate that Pab1 is also a part of the 3'-end RNA-processing complex and thus participates in the control of the poly(A) tail lengths during the polyadenylation reaction.
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Konopka, Catherine A., Melissa N. Locke, Pamela S. Gallagher, et al. "A yeast model for polyalanine-expansion aggregation and toxicity." Molecular Biology of the Cell 22, no. 12 (2011): 1971–84. http://dx.doi.org/10.1091/mbc.e11-01-0037.
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Nine human disorders result from the toxic accumulation and aggregation of proteins with expansions in their endogenous polyalanine (polyA) tracts. Given the prevalence of polyA tracts in eukaryotic proteomes, we wanted to understand the generality of polyA-expansion cytotoxicity by using yeast as a model organism. In our initial case, we expanded the polyA tract within the native yeast poly(Adenine)-binding protein Pab1 from 8A to 13A, 15A, 17A, and 20A. These expansions resulted in increasing formation of Pab1 inclusions, insolubility, and cytotoxicity that correlated with the length of the polyA expansion. Pab1 binds mRNA as part of its normal function, and disrupting RNA binding or altering cytoplasmic mRNA levels suppressed the cytotoxicity of 17A-expanded Pab1, indicating a requisite role for mRNA in Pab1 polyA-expansion toxicity. Surprisingly, neither manipulation suppressed the cytotoxicity of 20A-expanded Pab1. Thus longer expansions may have a different mechanism for toxicity. We think that this difference underscores the potential need to examine the cytotoxic mechanisms of both long and short expansions in models of expansion disorders.
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Anderson, J. T., M. R. Paddy, and M. S. Swanson. "PUB1 is a major nuclear and cytoplasmic polyadenylated RNA-binding protein in Saccharomyces cerevisiae." Molecular and Cellular Biology 13, no. 10 (1993): 6102–13. http://dx.doi.org/10.1128/mcb.13.10.6102-6113.1993.
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Proteins that directly associate with nuclear polyadenylated RNAs, or heterogeneous nuclear RNA-binding proteins (hnRNPs), and those that associate with cytoplasmic mRNAs, or mRNA-binding proteins (mRNPs), play important roles in regulating gene expression at the posttranscriptional level. Previous work with a variety of eukaryotic cells has demonstrated that hnRNPs are localized predominantly within the nucleus whereas mRNPs are cytoplasmic. While studying proteins associated with polyadenylated RNAs in Saccharomyces cerevisiae, we discovered an abundant polyuridylate-binding protein, PUB1, which appears to be both an hnRNP and an mRNP. PUB1 and PAB1, the polyadenylate tail-binding protein, are the two major proteins cross-linked by UV light to polyadenylated RNAs in vivo. The deduced primary structure of PUB1 indicates that it is a member of the ribonucleoprotein consensus sequence family of RNA-binding proteins and is structurally related to the human hnRNP M proteins. Even though the PUB1 protein is a major cellular polyadenylated RNA-binding protein, it is nonessential for cell growth. Indirect cellular immunofluorescence combined with digital image processing allowed a detailed comparison of the intracellular distributions of PUB1 and PAB1. While PAB1 is predominantly, and relatively uniformly, distributed within the cytoplasm, PUB1 is localized in a nonuniform pattern throughout both the nucleus and the cytoplasm. The cytoplasmic distribution of PUB1 is considerably more discontinuous than that of PAB1. Furthermore, sucrose gradient sedimentation analysis demonstrates that PAB1 cofractionates with polyribosomes whereas PUB1 does not. These results suggest that PUB1 is both an hnRNP and an mRNP and that it may be stably bound to a translationally inactive subpopulation of mRNAs within the cytoplasm.
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Anderson, J. T., M. R. Paddy, and M. S. Swanson. "PUB1 is a major nuclear and cytoplasmic polyadenylated RNA-binding protein in Saccharomyces cerevisiae." Molecular and Cellular Biology 13, no. 10 (1993): 6102–13. http://dx.doi.org/10.1128/mcb.13.10.6102.
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Proteins that directly associate with nuclear polyadenylated RNAs, or heterogeneous nuclear RNA-binding proteins (hnRNPs), and those that associate with cytoplasmic mRNAs, or mRNA-binding proteins (mRNPs), play important roles in regulating gene expression at the posttranscriptional level. Previous work with a variety of eukaryotic cells has demonstrated that hnRNPs are localized predominantly within the nucleus whereas mRNPs are cytoplasmic. While studying proteins associated with polyadenylated RNAs in Saccharomyces cerevisiae, we discovered an abundant polyuridylate-binding protein, PUB1, which appears to be both an hnRNP and an mRNP. PUB1 and PAB1, the polyadenylate tail-binding protein, are the two major proteins cross-linked by UV light to polyadenylated RNAs in vivo. The deduced primary structure of PUB1 indicates that it is a member of the ribonucleoprotein consensus sequence family of RNA-binding proteins and is structurally related to the human hnRNP M proteins. Even though the PUB1 protein is a major cellular polyadenylated RNA-binding protein, it is nonessential for cell growth. Indirect cellular immunofluorescence combined with digital image processing allowed a detailed comparison of the intracellular distributions of PUB1 and PAB1. While PAB1 is predominantly, and relatively uniformly, distributed within the cytoplasm, PUB1 is localized in a nonuniform pattern throughout both the nucleus and the cytoplasm. The cytoplasmic distribution of PUB1 is considerably more discontinuous than that of PAB1. Furthermore, sucrose gradient sedimentation analysis demonstrates that PAB1 cofractionates with polyribosomes whereas PUB1 does not. These results suggest that PUB1 is both an hnRNP and an mRNP and that it may be stably bound to a translationally inactive subpopulation of mRNAs within the cytoplasm.
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Cosson, Bertrand, Anne Couturier, Svetlana Chabelskaya, et al. "Poly(A)-Binding Protein Acts in Translation Termination via Eukaryotic Release Factor 3 Interaction and Does Not Influence [PSI+] Propagation." Molecular and Cellular Biology 22, no. 10 (2002): 3301–15. http://dx.doi.org/10.1128/mcb.22.10.3301-3315.2002.
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ABSTRACT Recent studies of translational control suggest that translation termination may not be simply the end of synthesizing a protein but rather be involved in modulating both the translation efficiency and stability of a given transcript. Using recombinant eukaryotic release factor 3 (eRF3) and cellular extracts, we have shown for Saccharomyces cerevisiae that yeast eRF3 and Pab1p can interact. This interaction, mediated by the N+M domain of eRF3 and amino acids 473 to 577 of Pab1p, was demonstrated to be direct by the two-hybrid approach. We confirmed that a genetic interaction exists between eRF3 and Pab1p and showed that Pab1p overexpression enhances the efficiency of termination in SUP35 (eRF3) mutant and [PSI +] cells. This effect requires the interaction of Pab1p with eRF3. These data further strengthen the possibility that Pab1p has a role in coupling translation termination events with initiation of translation. Several lines of evidence indicate that Pab1p does not influence [PSI +] propagation. First, “[PSI +]-no-more” mutations do not affect eRF3-Pab1p two-hybrid interaction. Second, overexpression of PAB1 does not cure the [PSI +] phenotype or solubilize detectable amounts of eRF3. Third, prion-curing properties of overexpressed HSP104p, which is required for formation and maintenance of [PSI +], were not modified by excess Pab1p.
Dissertations / Theses on the topic "Pab1"
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BRAMBILLA, MARCO. "Rewiring yeast nitrogen and mRNA metabolism for eliciting industrially relevant phenotypes. The Saccharomyces cerevisiae glutamate synthase (Glt1) and the poly(A) binding protein (Pab1) as case studies." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2018. http://hdl.handle.net/10281/198932.
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Le attuali produzioni di energia e materiali si basano soprattutto sullo sfruttamento di fonti fossili, ma la loro non rinnovabilità e la necessità di una maggiore sostenibilità ambientale han contribuito ad aumentare l’interesse verso risorse alternative. In questo contesto si collocano le bioraffinerie, che sono volte alla conversione di biomasse rinnovabili in prodotti commerciabili. Per implementare bioindustrie competitive, molte sfide devono essere superate, tra cui il miglioramento delle prestazioni dei microrganismi, cell factory, utilizzati nei processi produttivi. Tra tutte, il lievito Saccharomyces cerevisiae è la principale cell factory per la produzione di bioetanolo come biocarburante. L'obiettivo principale di questa tesi è stato di ingegnerizzare ceppi di S. cerevisiae per ottenere una cell factory con caratteristiche rilevanti a livello industriale, tra cui aumentate performances di crescita e un incremento della termotolleranza. In particolare, è stato studiato e quindi indotto un profondo rewiring cellulare, selezionando due proteine: la glutammato sintasi Glt1, un enzima del metabolismo centrale dell'azoto, CNM e la principale polyA binding protein Pab1, un fattore chiave nel metabolismo dell'mRNA. Gli effetti fisiologici della delezione e overespressione di GLT1 sono stati valutati coltivando S. cerevisiae con diverse fonti di azoto. I risultati mostrano che l'aggiunta di solfato d’ammonio, glutammato o glutammina, ma non la modulazione dell’espressione di GLT1, influenza notevolmente la crescita, il contenuto proteico, la vitalità e l'accumulo di specie reattive dell'ossigeno ROS. Questi dati evidenziano la plasticità del CNM di S. cerevisiae rispetto a variazioni ambientale e confermano la sua robustezza contro perturbazioni interne. Inoltre, sebbene la modulazione dell'espressione di GLT1 potrebbe non indurre una riprogrammare cellulare, la caratterizzazione fisiologica descritta puo essere utile per selezionare altri target pio promettenti per riprogrammazioni cellulari. L’induzione del rewiring cellulare è stato poi valutato selezionando Pab1 come target. A tal scopo, un ceppo recante la copia endogena di PAB1 è stato trasformato con una mutant library plasmidica di PAB1 e poi sottoposto a screening per isolare ceppi con maggior termotolleranza. I cloni isolati han mostrato, in drop test, una maggior crescita ad alte temperature e alte concentrazioni di etanolo. Tra tutte, la variante PAB1 S40.7 conferisce una maggiore termotolleranza tramite l’espressione dei soli primi 20 amminoacidi di Pab1. Tale fenotipo è stato anche confermato in bioreattore a 40°C. Una possibile ragione della maggior termotolleranza del ceppo S40.7 potrebbe essere collegata ad un minor accumulo di ROS rispetto al ceppo di controllo. Nel complesso, questi risultati han dimostrato che Pab1 è un promettente target per indurre fenotipi complessi con tratti migliorati, tra cui una maggiore termotolleranza. Infine, Pab1 è stata caratterizzata per determinare il ruolo dei suoi domini nel suo reclutamento all'interno di stress granules, SG, aggregati di mRNP non tradotti che si formano in condizioni stressanti. Questa caratterizzazione ha mostrato che l'associazione di Pab1 negli SG è principalmente dovuta agli RNA Recognition Motives RRM, il cui numero è importante per un efficiente reclutamento. Sebbene i domini P e C non partecipino direttamente all'associazione di Pab1 negli SG, la loro presenza rafforza o diminuisce, rispettivamente, la localizzazione di varianti sintetiche di Pab1 prive di almeno un RRM in questi aggregati. Questo lavoro suggerisce inoltre che i domini di Pab1 potrebbero essere sfruttati razionalmente per scopi di biologia sintetica. Nel complesso, i risultati di questa tesi evidenziano la difficoltà ma allo stesso tempo la capacità del rewiring cellulare nell’indurre fenotipi industrialmente rilevanti e, in particolare, Pab1 è emerso come target promettente per questo approccio.<br>Nowadays, for the production of energy and materials, our society mainly relies on fossil sources, but many concerns arise from their utilization, like greenhouse gases emission and non-renewability within the time of their consumption. Hence, biorefineries, which convert renewable biomasses into products and energy, could be a promising alternative. Despite some biorefineries are now at commercial scale, many challenges must be overcome to implement competitive bio-based industries, such as improving the performances of microorganisms, named cell factory, used during the production processes. Above all, the yeast Saccharomyces cerevisiae is the most prominent cell factory for producing bioethanol as biofuel. The main objective of this thesis was to engineer S. cerevisiae strains with biotechnological interesting traits, among which improved growth performances and increased thermotolerance. For this purpose, we investigated and applied a cellular rewiring, by selecting two targets: the glutamate synthase Glt1 that is an enzyme of the central nitrogen metabolism, CNM, and the main polyA binding protein Pab1, a master regulator of mRNA metabolism. Regarding GLT1, the physiological effects of its deletion and over-expression were assessed by growing yeasts in the presence of different nitrogen sources. Results showed that the supplementation of ammonium sulfate, glutamate or glutamine considerably affects growth, protein content, viability and reactive oxygen species, ROS, accumulation. Conversely, GLT1 modulation does not significantly influence these parameters. Overall, these data highlight the plasticity of the S. cerevisiae CNM in respect to the environment and confirm its robustness against internal perturbation. Moreover, even though the sole modulation of GLT1 expression might not reprogram the entire cell, the physiological characterization of this study might be helpful to guide the selection of other more promising candidate for the application of the rewiring approaches. Then, the induction of cellular reprogramming was assessed selecting Pab1. To this purpose, a strain carrying the unaltered PAB1 chromosomal allele was transformed with a PAB1 plasmid mutant library and then screened for isolating strains with high thermotolerance. The isolated clones showed growth improvement at both high temperatures and ethanol concentration by drop tests. Among all, the PAB1 S40.7 variant was further characterized because, strikingly, it dominantly confers higher thermotolerance by expressing just the first 20 amino acidic residues of Pab1. This improved phenotype was also confirmed in bioreactor at 40°C. Remarkably, the S40.7 strain accumulates less ROS compared to the control strain, thus possibly explaining its increased thermotolerance. Overall, these results demonstrated that Pab1 is a powerful candidate to evoke complex phenotypes with improved traits, among which higher thermotolerance. Finally, Pab1 was characterized to uncover the role of its domains in the recruitment of the protein within stress granules, aggregates of untranslated mRNPs that form during stressful conditions. This characterization shows that Pab1 association into these aggregates relies mainly on RNA recognition motives, RRM, whose number is important for an efficient recruitment. Although the P and C domains do not directly participate in Pab1 association to stress granules, their presence strengthens or decreases, respectively, the distribution of synthetic Pab1 variants lacking at least one RRM into these aggregates. As additional outcome, this part of the work is suggesting that Pab1 domains might be rationally exploited for synthetic biology purposes. Overall, the results of this thesis highlight and confirm the difficulty but at the same time the power of cellular rewiring in evoking industrially relevant phenotype. Pab1 is undoubtedly emerging as a pivotal element that deserves more attention for future strain design and tailoring.
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Grenier, St-Sauveur Valérie. "Caractérisation et fonction de la protéine Nab2 chez Schizosaccharomyces pombe." Mémoire, Université de Sherbrooke, 2014. http://savoirs.usherbrooke.ca/handle/11143/105.
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L’étude qui vous est présentée dans ce mémoire est réalisée chez l’organisme Schizosaccharomyces pombe et elle porte sur la caractérisation de la protéine Nab2, une protéine liant les queues poly(A) ainsi que sur son implication dans la régulation génique. Tout d’abord, il faut savoir qu’il existe quatre modes de régulation (transcriptionnel, post-transcriptionnel, traductionnel et post-traductionnel) dans lesquels interviennent différents types de protéines, en particulier des protéines liant les queues poly(A) des ARN, aussi connues sous le nom de PABPs. Ces protéines reconnaissent l’ARN à l’aide de différents domaines de liaison et elles se subdivisent en deux catégories : les PABPs nucléaires ou cytoplasmiques, représentées respectivement par PABPN1 et PABPC1 chez les mammifères. Comprendre la fonction des PABPs revêt un intérêt particulier puisqu’elles sont impliquées à différents stades de la régulation génique. Des maladies ont aussi été associées à deux PABPs nucléaires humaines, PABPN1 et ZC3H14, mais aucune association entre leur fonction réciproque et la maladie n’a pu être établie. Une des façons de comprendre leur rôle est d’étudier celui de leurs orthologues respectifs. Chez la levure à fission, un orthologue de PABPN1 a été caractérisé et il s’agit de Pab2. S. pombe possède cependant une seconde PABP nucléaire, Nab2, qui est caractérisée dans ces travaux. Des méthodes in vivo et in vitro ont été utilisées afin de confirmer le statut de la protéine, à savoir qu’il s’agit bel et bien d’une PABP nucléaire et que celle-ci est non essentielle. L’identification de partenaires protéiques de Nab2 par spectrométrie de masse a aussi permis de relier Nab2 avec des processus de régulation génique tels que l’épissage et la dégradation. Puisque Pab2 est aussi associée à des fonctions en lien avec la dégradation, il est possible de faire un parallèle entre ces deux protéines et de supposer qu’elles interagissent ensemble. La deuxième partie de ces travaux porte donc sur l’étude de la relation fonctionnelle entre Nab2 et Pab2 et elle a permis de montrer un mécanisme de régulation opportuniste basé sur la liaison de la cible ARN par l’une ou l’autre de ces PABPs. En effet, l’étude de la régulation du gène modèle RPL30-2 indique que Nab2 et Pab2 ont des rôles opposés puisqu’ils sont respectivement des régulateurs positifs et négatifs de l’expression de son transcrit.
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Grenier, St-Sauveur Val??rie. "Caract??risation et fonction de la prot??ine Nab2 chez Schizosaccharomyces pombe." Mémoire, Universit?? de Sherbrooke, 2014. http://savoirs.usherbrooke.ca/handle/11143/105.
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L?????tude qui vous est pr??sent??e dans ce m??moire est r??alis??e chez l???organisme Schizosaccharomyces pombe et elle porte sur la caract??risation de la prot??ine Nab2, une prot??ine liant les queues poly(A) ainsi que sur son implication dans la r??gulation g??nique. Tout d???abord, il faut savoir qu???il existe quatre modes de r??gulation (transcriptionnel, post-transcriptionnel, traductionnel et post-traductionnel) dans lesquels interviennent diff??rents types de prot??ines, en particulier des prot??ines liant les queues poly(A) des ARN, aussi connues sous le nom de PABPs. Ces prot??ines reconnaissent l???ARN ?? l???aide de diff??rents domaines de liaison et elles se subdivisent en deux cat??gories : les PABPs nucl??aires ou cytoplasmiques, repr??sent??es respectivement par PABPN1 et PABPC1 chez les mammif??res. Comprendre la fonction des PABPs rev??t un int??r??t particulier puisqu???elles sont impliqu??es ?? diff??rents stades de la r??gulation g??nique. Des maladies ont aussi ??t?? associ??es ?? deux PABPs nucl??aires humaines, PABPN1 et ZC3H14, mais aucune association entre leur fonction r??ciproque et la maladie n???a pu ??tre ??tablie. Une des fa??ons de comprendre leur r??le est d?????tudier celui de leurs orthologues respectifs. Chez la levure ?? fission, un orthologue de PABPN1 a ??t?? caract??ris?? et il s???agit de Pab2. S. pombe poss??de cependant une seconde PABP nucl??aire, Nab2, qui est caract??ris??e dans ces travaux. Des m??thodes in vivo et in vitro ont ??t?? utilis??es afin de confirmer le statut de la prot??ine, ?? savoir qu???il s???agit bel et bien d???une PABP nucl??aire et que celle-ci est non essentielle. L???identification de partenaires prot??iques de Nab2 par spectrom??trie de masse a aussi permis de relier Nab2 avec des processus de r??gulation g??nique tels que l?????pissage et la d??gradation. Puisque Pab2 est aussi associ??e ?? des fonctions en lien avec la d??gradation, il est possible de faire un parall??le entre ces deux prot??ines et de supposer qu???elles interagissent ensemble. La deuxi??me partie de ces travaux porte donc sur l?????tude de la relation fonctionnelle entre Nab2 et Pab2 et elle a permis de montrer un m??canisme de r??gulation opportuniste bas?? sur la liaison de la cible ARN par l???une ou l???autre de ces PABPs. En effet, l?????tude de la r??gulation du g??ne mod??le RPL30-2 indique que Nab2 et Pab2 ont des r??les oppos??s puisqu???ils sont respectivement des r??gulateurs positifs et n??gatifs de l???expression de son transcrit.
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Penheiter, Kristi L. "Functional characterization of the Paf1 complex in Saccharomyces cerevisiae by identification of Paf1 target genes /." Connect to full text via ProQuest. IP filtered, 2005.
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Thesis (Ph.D. in Molecular Biology) -- University of Colorado at Denver and Health Sciences Center, 2005.<br>Typescript. Includes bibliographical references (leaves 126-149). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;
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de, Carli da Costa Lima Tamara. "Caracterização funcional de homólogos à proteína de ligação a Cauda Poli-A (PABP) de Leishmania major." Universidade Federal de Pernambuco, 2007. https://repositorio.ufpe.br/handle/123456789/6362.
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Previous issue date: 2007<br>A proteína de ligação à cauda poli-A (PABP) se liga a seqüência de poli
adenosinas presente na extremidade 3 do mRNA e possui uma
multiplicidade de funções dentro da célula. Dentre as funções atribuídas
a PABP destaca-se sua participação em eventos da síntese de proteínas,
tais como iniciação e terminação da tradução e reciclagem dos
ribossomos, e seu envolvimento no transporte de alguns mRNAs do
núcleo para o citoplasma. O objetivo deste trabalho é caracterizar
homólogos de proteínas relevantes à iniciação da tradução em
tripanosomatídeos. O estudo da PABP teve início com a busca em bancos
de dados de L. major de possíveis homólogos, onde foi encontrado tanto
o homólogo previamente descrito (LmPABP1) como mais dois homólogos
(LmPABP2 e LmPABP3), sendo que o último não possui ortólogos em
Trypanosoma. Os três genes foram amplificados, clonados, as proteínas
expressas e utilizadas para produção de soro policlonal. Em seguida foi
realizada a quantificação dos níveis intracelulares das três proteínas em
extratos da forma promastigota de L. major sendo a LmPABP2 a mais
abundante, a LmPABP3 em níveis intermediários e a LmPABP1 a menos
abundante. No entanto, as LmPABP2-3 são detectadas com uma única
forma, enquanto que a LmPABP1 está presente em duas isoformas,
provavelmente uma delas devido a fosforilação. Análises da expressão
durante o ciclo evolutivo do parasita mostrou que as três proteínas
encontravam-se presentes em todas as formas evolutivas, porém a
LmPABP3 mostrou um decréscimo na fase estacionária de crescimento e
a LmPABP1 apresenta-se fosforilada apenas nas primeiras horas após o
repique. Experimentos de localização subcelular indicam que a LmPABP1
está presente na fração citoplasmática, enquanto que LmPABP2-3 estão
presentes tanto na fração citoplasmática quanto na nuclear, porém com
predominância da fração nuclear. Estudos adicionais precisam ser feitos
pra entender como estas proteínas diferem funcionalmente e quais seus
papéis na síntese protéica
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Siddiqui, Nadeem. "Structure and function of the PABC domain." Thesis, McGill University, 2006. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=102727.
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The poly (A)-binding protein (PABP) is an essential protein found in all eukaryotes and functions in mRNA metabolic and translational processes. Structurally, PABP consists of two distinct regions. The N-terminal half contains four RNA recognition motifs that bind to the poly (A)-tail of mRNA, while the C-terminal segment contains a unique peptide binding module referred to as the PABC domain. The function of this domain in PABP is to recruit proteins containing a very specific 'PAM-2' motif to the mRNP complex. Unique to metazoans, a PABC domain is also found in the hyperplastic discs tumor suppressor (HYD), which is an E3 ubiquitin ligase.<br>This thesis completes a structural investigation of PABC domains from various species by nuclear magnetic resonance spectroscopy. In particular, we report the solution structure of PABC from the parasite Trypanosoma cruzi and plant Triticum aevestium PABP. Both domains consist of five alpha-helices which fold into a structure highly comparable to the human PABC domain from PABP and HYD. All four PABCs interact with a similar PAM-2 sequence and show comparable peptide binding surfaces. The human PABC-PAM-2 complex structure displays a PAM-2 peptide interacting with specific residues within the domain. Sequence analyses and peptide surface mapping studies show that these residues are highly conserved, which indicates an analogous mechanism of peptide recognition throughout animal, parasite, and plant species. An exception to these observations was found in the PABC domain from Saccharomyces cerevisiae PABP. Yeast PABC recognizes a variation of the typical PAM-2 motif but mediates its interaction through a similar mechanism as human PABC.<br>The PAM-2 motif encloses a signature sequence which was used to successfully identify new interacting partners for the PABC domain via a bioinformatics screen. In mammalian systems, the identified proteins are implicated in either RNA metabolic, translational, or ubiquitin associated functions. This thesis concludes with a model illustrating a unique cross-talk between major gene expression pathways mediated by the PABC domain and its binding partners.
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Tsui, Hoyee. "The role of p21-activated kinase 1 (Pak1) in the heart." Thesis, University of Manchester, 2015. https://www.research.manchester.ac.uk/portal/en/theses/the-role-of-p21activated-kinase-1-pak1-in-the-heart(8c34d7bc-a2aa-4ae0-a197-91ed905212f5).html.
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Heart failure is associated with a high mortality rate and is one of the most prevalent diseases worldwide whereby susceptibility increases with age. The development of heart failure occurs over an extensive period of time in which arrhythmias and hypertrophy are both very prevalent manifestations throughout this progression. Arrhythmias are defined as an irregular rhythm originating from intracellular calcium dysregulation, which can be fatal. Cardiac hypertrophy is a compensatory condition induced by increased workload involving augmented cardiomyocyte growth accompanied by myocardial remodelling. However, under prolonged periods of increased stress this compensatory mechanism can lead to cardiac dysfunction. The current treatments for heart failure are mainly aimed at relieving symptoms or itself possess proarrhythmic ability. Therefore it is fundamental to elucidate the pathways involved in arrhythmias and hypertrophy for the development of more effective treatment. p21 activated protein kinase (Pak1) is a novel gene involved in the regulation of cardiac function, however, the mechanisms involved remain inconclusive. This study has demonstrated Pak1 to be both antiarrhythmic and antihypertrophic, emphasizing Pak1 as a credible therapeutic target for simultaneously treating both manifestations. The antiarrhythmic properties of Pak1 were demonstrated through cardiomyocyte-specific Pak1 knockout (Pak1cko) mouse model which underwent Isoproterenol (ISO) stimulation for 2 weeks. Compared with ISO treated control group, the Pak1cko group had increased calcium irregularities and particularly a prolongation in sarcoplasmic reticulum (SR) calcium refill. The absence of Pak1 abrogated the transcriptional up-regulation of sarcoplasmic reticulum calcium ATPase 2a (SERCA2a) under stressed conditions. Further analysis in neonatal rat cardiomyocytes (NRCMs) revealed this regulation to be through activation of the transcription factor, SRF. The antihypertrophic effects of Pak1 were further illustrated through cardiomyocyte-specific overexpressed constitutively-active Pak1 (Pak1cTG) mice which were subjected to transverse aortic constriction (TAC) for 3 weeks. Compared to TAC control group, Pak1cTG mice had improved cardiac performance accompanied with diminished fibrosis. Further analysis led to the discovery of a novel antihypertrophic pathway of Pak1 involving positive regulation of the E3ligase, Fbxo32 through activation of Smad3. This pathway is vital in the prevention of calcineurin (PP2B) accretion. Berberine administration in TAC treated mice corroborated that Fbxo32 up-regulation is sufficient in the prevention of hypertrophy. In conclusion, my study has demonstrated that Pak1 conveys antiarrhythmic influence through the up-regulation of SERCA2a. In the prevention of pathological hypertrophy, Pak1 inhibits PP2B through positive regulation of Fbxo32. Overall, my thesis has advanced the knowledge about cardioprotective pathways initiated by Pak1 under stressed conditions, presenting Pak1 as a promising therapeutic target.
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Ganem, Carine. "Implication de Cak1 (CDK activating kinase) dans le processus de transcription par l'ARN polymérase II chez Saccharomyces cerevisiae." Paris 11, 2002. http://www.theses.fr/2002PA112180.
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Chez la levure S. Cerevisiae, les rôles physiologiques connus de la protéine essentielle Cak1 (kinase activatrice des kinases cycline-dépendantes (CDK)) sont de phosphoryler Cdc28, régulateur majeur du cycle cellulaire, ainsi que Kin28, kinase du domaine carboxy-terminal de la plus grosse sous-unité de l'ARN polymérase II. Des mutants conditionnels de cakI semblent affectés dans le contrôle du cycle cellulaire, mais aussi dans la formation de la paroi des spores, la différentiation pseudohyphale, ainsi que dans la réponse au stress osmotique. Dans ce travail, nous avons montré que Cak1 est impliquée dans le processus de transcription non seulement via la phosphorylation de Kin28, mais également via le complexe transcriptionnel multimérique Paf1 et via la protéine Ssu72. Une étude plus poussée de Ssu72 nous a permis de dévoiler sa nature de protéine phosphatase, et les liens étroits, tant génétiques que moléculaires, qu'elle tisse avec plusieurs autres protéines impliquées dans l'expression des gènes, telles que Sua7 (TFIIB), Kin28, Pafl, Ctr9 ou Fcp1. La multiplicité des effets des mutations dans le gène CAK1 pourrait ainsi s'expliquer par un rôle de modulateur de l'activité de transcription<br>In the budding yeast S. Cerevisae, the known physiological roles of the essential protein Cak1 (Cyclin-dependant kinases (CDKs) Activating Kinase) are to phosphorylate Cdc28, the main regulator of progression through cell cycle, and Kin28, the carboxy-terminal domain (CTD) kinase of the largest RNA polymerase II subunit. Conditionnal mutants of cakl appear affected in cell cycle control, but also in spore-wall morphogenesis, pseudohyphal differentiation, and response to high osmolarity. In this work, we show that Cak1 is implicated in transcription process not only through Kin28 phosphorylation, but also through the multimeric transcriptional complex Paf1 and the Ssu72 protein. A more extensive study of Ssu72 enabled us to reveal its phosphatase activity, as well as its genetic and molecular interactions with several other proteins involved in gene expression, such as Sua7 (TFIIB), Kin28, Paf1, Ctr9 or Fcp1. Thus, the multiple effects of cakl mutations could be explained by the role of Cak1 as a modulator of transcriptional activity
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Angathevar, Veluchamy Raaja Raajan. "Structure and activity of Pseudomonas aeruginosa PAO1 biofilms." Thesis, Montana State University, 2006. http://etd.lib.montana.edu/etd/2006/angathevarveluchamy/AngathevarVeluchamyR0506.pdf.
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Roy, Guylaine. "Characterization of PABP-interacting proteins 1 and 2." Thesis, McGill University, 2003. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=84317.
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The 3' poly(A) tail of eukaryotic mRNAs and the poly(A) binding protein (PABP) play key roles in the regulation of translation. Recently, our group identified two human PABP-interacting proteins (Paip), Paip1 and Paip2, which stimulate and repress translation, respectively. Paip2 also inhibits the binding of PABP to the poly(A) tail and competes with Paip1 for binding to PABP. My research project was divided into two parts to allow me to gain a greater understanding of the roles of these two PABP-interacting proteins. First, in order to study the mechanism of interaction of Paip1 and PABP, their binding sites were mapped by Far Western and GST pull-downs experiments. The Paip1-PABP interaction involves two distinct binding regions in each protein. The PABP interacting motif-1 (PAM1) of Paip1 is rich in acidic amino acids and is located in the C-terminus (a.a. 440--479). PAM1 interacts with the RNA recognition motifs (RRMs) 1 and 2 of PABP. PAM2 consists of a 15 amino acid stretch residing in the N-terminus of Paip1 (a.a. 123--137) and interacts with the C-terminal domain of PABP. In addition, the stoichiometry and the kinetic and thermodynamic constants for the Paip1-PABP interaction were determined using a Surface Plasmon Resonance (SPR)-biosensor. Paip1 interacts with PABP with an apparent KD of 1.9 nM and with a 1:1 stoichiometry. In the second part of my thesis research, the Drosophila Paip2 (dPaip2) was isolated and characterized in order to ascertain a biological role for Paip2. dPaip2 was found to be the bona fide homologue of human Paip2 since it interacts with the Drosophila PABP (dPABP) via two independent binding sites, interferes with the ability of dPABP to bind to poly(A), and represses translation. Ectopic overexpression of dPaip2 in wings resulted in a size reduction phenotype, which was due to a decrease in the cell number but not to a difference in cell size. Clones of cells overexpressing dPaip2 in wing discs also contai
Books on the topic "Pab1"
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d1920-, Kim T'ae-gil. Chagŭn pabo wa k'ŭn pabo: Kim T'ae-gil eseijip. Chayu Munhaksa, 1986.
Find full textBook chapters on the topic "Pab1"
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Bährle-Rapp, Marina. "PABA." In Springer Lexikon Kosmetik und Körperpflege. Springer Berlin Heidelberg, 2007. http://dx.doi.org/10.1007/978-3-540-71095-0_7324.
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Gooch, Jan W. "PABM." In Encyclopedic Dictionary of Polymers. Springer New York, 2011. http://dx.doi.org/10.1007/978-1-4419-6247-8_8352.
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Braun-Falco, Markus, Henry J. Mankin, Sharon L. Wenger, et al. "PAH1." In Encyclopedia of Molecular Mechanisms of Disease. Springer Berlin Heidelberg, 2009. http://dx.doi.org/10.1007/978-3-540-29676-8_9500.
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Gressner, A. M., and O. A. Gressner. "PABA-Test." In Lexikon der Medizinischen Laboratoriumsdiagnostik. Springer Berlin Heidelberg, 2017. http://dx.doi.org/10.1007/978-3-662-49054-9_2333-1.
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Gressner, A. M., and O. A. Gressner. "PABA-Test." In Springer Reference Medizin. Springer Berlin Heidelberg, 2019. http://dx.doi.org/10.1007/978-3-662-48986-4_2333.
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Bährle-Rapp, Marina. "Allantoin Paba." In Springer Lexikon Kosmetik und Körperpflege. Springer Berlin Heidelberg, 2007. http://dx.doi.org/10.1007/978-3-540-71095-0_339.
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Bährle-Rapp, Marina. "PEG-25 PABA." In Springer Lexikon Kosmetik und Körperpflege. Springer Berlin Heidelberg, 2007. http://dx.doi.org/10.1007/978-3-540-71095-0_7652.
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Bashford, David. "Polyamide 11 (PA11)." In Thermoplastics. Springer Netherlands, 1997. http://dx.doi.org/10.1007/978-94-009-1531-2_49.
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Bährle-Rapp, Marina. "Ethyl Dihydroxypropyl PABA." In Springer Lexikon Kosmetik und Körperpflege. Springer Berlin Heidelberg, 2007. http://dx.doi.org/10.1007/978-3-540-71095-0_3713.
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Bährle-Rapp, Marina. "Ethylhexyl Dimethyl PABA." In Springer Lexikon Kosmetik und Körperpflege. Springer Berlin Heidelberg, 2007. http://dx.doi.org/10.1007/978-3-540-71095-0_3745.
Full textConference papers on the topic "Pab1"
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Singhal, Rekha, and Dheeraj Chahal. "PABS 2015." In ICPE'15: ACM/SPEC International Conference on Performance Engineering. ACM, 2015. http://dx.doi.org/10.1145/2668930.2688199.
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Singhal, Rekha, and Dheeraj Chahal. "Session details: PABS'16." In ICPE'16: ACM/SPEC International Conference on Performance Engineering. ACM, 2016. http://dx.doi.org/10.1145/3257768.
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Fontinele, Alexandre Cardoso, Iallen Gábio de Sousa Santos, Jurandir Cavalcante Lacerda Júnior, André Castelo Branco Soares, Adolfo Da Visitação Tregeira Cartaxo, and Divanilson Rodrigo de Sousa Campelo. "Novo Algoritmo para Atribuição de Potência por Circuito em Redes Ópticas Elásticas." In Simpósio Brasileiro de Redes de Computadores e Sistemas Distribuídos. Sociedade Brasileira de Computação, 2020. http://dx.doi.org/10.5753/sbrc.2020.12309.
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Este artigo apresenta um estudo em redes ópticas elásticas considerando o problema de Power, Routing, Modulation Level and Spectrum Assignment (PRMLSA). O presente estudo se concentra na atribuição de potência por circuito. Nesse contexto, o algoritmo de Power Assignment by Binary Search (PABS), um novo algoritmo que realiza atribuição de potência por circuito de forma adaptativa, é proposto. O algoritmo PABS é comparado com outros algoritmos presentes na literatura em termos de probabilidade de bloqueio de banda. O algoritmo PABS consegue uma redução na probabilidade de bloqueio de banda de pelo menos 39% na topologia NSFNet e 42% na topologia Cost239.
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Knaack, J., B. Ramsauer, RL Schild, T. Groten, M. Abou-Dakn, and U. Schäfer-Graf. "PABS: Pregnancy after bariatric surgery." In 64. Kongress der Deutschen Gesellschaft für Gynäkologie und Geburtshilfe e. V. Georg Thieme Verlag, 2022. http://dx.doi.org/10.1055/s-0042-1756901.
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Monroe, D. M., G. B. Sherrill, and H. R. Roberts. "A SIMPLE AND CONVENIENT FLUORESCENT ASSAY FOR MONITORING ACTIVATION OF COAGULATION FACTORS IX, X, AND PROTHROMBIN USING p-AMINO-BENZAMIDINE." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643563.
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Blood coagulation consists of a series of protein-protein-cofactor interactions that involve zymogen coagulation factors being activated to trypsin-like serine proteases, ultimately leading to conversion of prothrombin to thrombin. As described by Evans et al. ((1982) J. Biol. Chea. 257, 3014), the fluorescent compound p-aminobenzamidine (pAB) increases its quantum yield when bound to the active site of thrombin (but not when pAB is reacted with coagulation zymogens). We have devised a simple procedure to monitor activation of purified coagulation factors IX, X, and prothrombin using pAB. Human factor X with 150 μM pAB was activated with purified factor X activating protein from Russell’s Viper venom (RVVX). Fluorescence emission was monitored at 376nm with excitation at 336nm. Portions were removed and activation was measured by clotting assay, by assay using tosyl-Gly-Pro-Arg-p-nitroanilide (tosGlyProArgNA), and by polyacrylamide gel electrophoresis; all of which indicated that factor X activation followed a time course identical to the time course described by the pAB fluorescence increase. Factor IX was activated by factor XIa in the presence of pAB; portions were analyzed for clotting activity. Prothrombin was activated to (meizo)thrombin by Ecarin and activity was measured using tosGly-ProArgNA. In both cases the increase in pAB fluorescence was coincident with the time course described by activity assay. The pAB fluorescence assay has several advantages including: 1) activation can be easily and continuously monitored, 2) the extent of modification can be determined by inspection of the progress curve, and 3) the procedure can be used with a wide range of zymogen concentrations. Additionally, pAB can be used to follow activation of modified (pro)enzymes. For instance, activation of bovine des(γcarboxyglutamic acid)factor X by RVVX was found to be 100-fold slower than activation of unmodified bovine factor X. Our results suggest that following p-aminobenzamidine fluorescence is a useful and convenient procedure for monitoring activation of blood coagulation factors IX, X, and prothrombin.
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Harikrishna Narasimhan. "Parallel artificial bee colony (PABC) algorithm." In 2009 World Congress on Nature & Biologically Inspired Computing (NaBIC). IEEE, 2009. http://dx.doi.org/10.1109/nabic.2009.5393726.
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Shi, Xiang, Lin Wang, Fa Zhang, Kai Zheng, and Zhiyong Liu. "PABO: Congestion mitigation via packet bounce." In ICC 2017 - 2017 IEEE International Conference on Communications. IEEE, 2017. http://dx.doi.org/10.1109/icc.2017.7996995.
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Shattil, S. J., J. A. Hoxie, M. Cunningham, C. S. Abrahms, J. O’Brien, and Z. Budzynski. "DETECTION OF ACTIVATED PLATELETS IN WHOLE BLOOD BY FLOW CYTOMETRY." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643830.
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Platelets may become activated in a number of clinical disorders and participate in thrombus formation. We have developed a direct test for activated platelets in whole blood that utilizes dual-color flow cytometry and requires no washing steps. Platelets were distinguished from erythrocytes and white blood cells in the flow cytometer by labeling the platelets with biotin-AP1, an antibody specific for membrane glycoprotein lb, and analyzing the cells for phycoerythrin-streptavidin fluorescence. Membrane surface changes resulting from platelet activation were detected with three different FITC-labeled monoclonal antibodies: 1) PAC1, an antibody specific for the fibrinogen receptor on activated platelets; 2) 9F9, which binds to the D-domain of fibrinogen and detects platelet-bound fibrinogen; and 3) S12, which binds to an alpha-granule membrane protein that associates with the platelet surface during secretion. Unstimulated platelets demonstrated no PAC1, 9F9, or S12-specific fluorescence, indicating that they did not bind these antibodies. Upon stimulation with agonists, however, the platelets demonstrated a dose-dependent increase in FITC-fluorescence. The binding of 9F9 to activated platelets required fibrinogen. Low concentrations of ADP and epinephrine, which induce fibrinogen receptors but little secretion, stimulated near-maximal PAC1 or 9F9 binding but little S12 binding. On the other hand, a concentration of phorbol myristate acetate that evokes full platelet aggregation and secretion induced maximal binding of all three antibodies. When blood samples containing activated and non-activated platelets were mixed, as few as 0.8% activated platelets could be detected by this technique. There was a direct correlation between ADP-induced FITC-PAC1 binding and binding determined in a conventional 125I-PAC1 binding assay (r = 0.99; p < 0.001). These studies demonstrate that activated platelets can be reliably detected in whole blood using activation-dependent monoclonal antibodies and flow cytometry. This method may be useful to assess the degree of platelet activation and the efficacy platelet inhibitor therapy in thrombotic disorders.
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Chang, Ed, and Tat-Seng Chua. "Session details: Panel - PA1." In MM '10: ACM Multimedia Conference. ACM, 2010. http://dx.doi.org/10.1145/3252111.
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Rabelo, Jacilane, Davi Viana, Gleison Santos, and Tayana Conte. "Usando PABC-Pattern para Codificar o Conhecimento: Um Estudo Experimental." In XIII Simpósio Brasileiro de Qualidade de Software. Sociedade Brasileira de Computação - SBC, 2014. http://dx.doi.org/10.5753/sbqs.2014.15240.
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Gerência do conhecimento tem um papel fundamental no desenvolvimento de software. A coleta de lições aprendidas é uma prática comum em organizações de software para gerenciar e transferir o conhecimento. A codificação do conhecimento pode facilitar o registro das lições aprendidas para sua consulta posterior. Este trabalho apresenta um estudo qualitativo da PABC- Pattern (Problema, Ação, Benefício e Contexto – Padrão), uma nova abordagem para codificar e compartilhar o conhecimento na forma de lições aprendidas. Os resultados mostraram que há uma facilidade de uso nos campos da abordagem Adicionalmente, foi feita uma comparação do esquema proposto pela PABC-Pattern com os esquemas de outras abordagens baseadas em padrão.
Reports on the topic "Pab1"
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Elroy-Stein, Orna, and Dmitry Belostotsky. Mechanism of Internal Initiation of Translation in Plants. United States Department of Agriculture, 2010. http://dx.doi.org/10.32747/2010.7696518.bard.
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Original objectives Elucidation of PABP's role in crTMV148 IRES function in-vitro using wheat germ extract and krebs-2 cells extract. Fully achieved. Elucidation of PABP's role in crTMV148 IRES function in-vivo in Arabidopsis. Characterization of the physical interactions of PABP and other potential ITAFs with crTMV148 IRES. Partly achieved. To conduct search for additional ITAFs using different approaches and evaluate the candidates. Partly achieved. Background of the topic The power of internal translation via the activity of internal ribosomal entry site (IRES) elements allow coordinated synthesis of multiple gene products from a single transcription unit, and thereby enables to bypass the need for sequential transformation with multiple independent transgenes. The key goal of this project was to identify and analyze the IRES-trans-acting factors (ITAFs) that mediate the activity of a crucifer-infecting tobamovirus (crTMV148) IRES. The remarkable conservation of the IRES activity across the phylogenetic spectrum (yeast, plants and animals) strongly suggests that key ITAFs that mediate its activity are themselves highly conserved. Thus, crTMV148 IRES offers opportunity for elucidation of the fundamental mechanisms underlying internal translation in higher plants in order to enable its rational manipulation for the purpose of agricultural biotechnology. Major conclusions and achievements. - CrTMV IRES requires PABP for maximal activity. This conclusion was achieved by PABP depletion and reconstitution of wheat germ- and Krebs2-derived in-vitro translation assays using Arabidopsis-derived PABP2, 3, 5, 8 and yeast Pab1p. - Mutations in the internal polypurine tract of the IRES decrease the high-affinity binding of all phylogenetically divergent PABPs derived from Arabidopsis and yeast in electro mobility gel shift assays. - Mutations in the internal polypurine tract decrease IRES activity in-vivo. - The 3'-poly(A) tail enhances crTMV148 IRES activity more efficiently in the absence of 5'-methylated cap. - In-vivo assembled RNPs containing proteins specifically associated with the IRES were purified from HEK293 cells using the RNA Affinity in Tandem (RAT) approach followed by their identification by mass spectroscopy. - This study yielded a list of potential protein candidates that may serve as ITAFs of crTMV148 IRES activity, among them are a/b tubulin, a/g actin, GAPDH, enolase 1, ribonuclease/angiogenin inhibitor 1, 26S proteasome subunit p45, rpSA, eEF1Bδ, and proteasome b5 subunit. Implications, both scientific and agriculture. The fact that the 3'-poly(A) tail enhances crTMV148 IRES activity more efficiently in the absence of 5'-methylated cap suggests a potential joint interaction between PABP, the IRES sequence and the 3'-poly(A). This has an important scientific implication related to IRES function in general.
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Schrot, John F., and J. R. Thomas. NP Pab Methodologies (NPPABINT). Defense Technical Information Center, 1991. http://dx.doi.org/10.21236/ada249866.
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Reid, Gary B. UTC-PAB Normative Database (PABNORM). Defense Technical Information Center, 1990. http://dx.doi.org/10.21236/ada242456.
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Spadaccini, Chris, Eric Duoss, Maxim Shusteff, Angela Tooker, and Razi Haque. Swab Testing Results- HP PA11 Rev 8 Summary. Office of Scientific and Technical Information (OSTI), 2020. http://dx.doi.org/10.2172/1631515.
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Spadaccini, Chris, Eric Duoss, Maxim Shusteff, Angela Tooker, and Razi Haque. Swab Tensile Testing Results and Procedures - PA11 Rev. 10 Swabs. Office of Scientific and Technical Information (OSTI), 2020. http://dx.doi.org/10.2172/1630403.
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Caron, D. S., E. Browne, and E. B. Norman. PABS: A Computer Program to Normalize Emission Probabilities and Calculate Realistic Uncertainties. Office of Scientific and Technical Information (OSTI), 2009. http://dx.doi.org/10.2172/973606.
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Hui, Liang, Liu Jie, Huang Hui, et al. Evaluation of PAX1 Methylation Triage as the Primary Screening Method in the Application of Early Diagnosis for Cervical Cancer. "Prof. Marin Drinov" Publishing House of Bulgarian Academy of Sciences, 2019. http://dx.doi.org/10.7546/crabs.2019.06.17.
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Olsen, Matthew. Investigation of Speech Samples from Typically Developing Preschool Age Children: A Comparison of Single Words and Imitated Sentences Elicited with the PABA-E. Portland State University Library, 2000. http://dx.doi.org/10.15760/etd.434.
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Coplin, David L., Shulamit Manulis, and Isaac Barash. roles Hrp-dependent effector proteins and hrp gene regulation as determinants of virulence and host-specificity in Erwinia stewartii and E. herbicola pvs. gypsophilae and betae. United States Department of Agriculture, 2005. http://dx.doi.org/10.32747/2005.7587216.bard.
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Gram-negative plant pathogenic bacteria employ specialized type-III secretion systems (TTSS) to deliver an arsenal of pathogenicity proteins directly into host cells. These secretion systems are encoded by hrp genes (for hypersensitive response and pathogenicity) and the effector proteins by so-called dsp or avr genes. The functions of effectors are to enable bacterial multiplication by damaging host cells and/or by blocking host defenses. We characterized essential hrp gene clusters in the Stewart's Wilt of maize pathogen, Pantoea stewartii subsp. stewartii (Pnss; formerly Erwinia stewartii) and the gall-forming bacterium, Pantoea agglomerans (formerly Erwinia herbicola) pvs. gypsophilae (Pag) and betae (Pab). We proposed that the virulence and host specificity of these pathogens is a function of a) the perception of specific host signals resulting in bacterial hrp gene expression and b) the action of specialized signal proteins (i.e. Hrp effectors) delivered into the plant cell. The specific objectives of the proposal were: 1) How is the expression of the hrp and effector genes regulated in response to host cell contact and the apoplastic environment? 2) What additional effector proteins are involved in pathogenicity? 3) Do the presently known Pantoea effector proteins enter host cells? 4) What host proteins interact with these effectors? We characterized the components of the hrp regulatory cascade (HrpXY ->7 HrpS ->7 HrpL ->7 hrp promoters), showed that they are conserved in both Pnss and Fag, and discovered that the regulation of the hrpS promoter (hrpSp) may be a key point in integrating apoplastic signals. We also analyzed the promoters recognized by HrpL and demonstrated the relationship between their composition and efficiency. Moreover, we showed that promoter strength can influence disease expression. In Pnss, we found that the HrpXY two-component signal system may sense the metabolic status of the bacterium and is required for full hrp gene expression in planta. In both species, acyl-homoserine lactone-mediated quorum sensing may also regulate epiphytic fitness and/or pathogenicity. A common Hrp effector protein, DspE/WtsE, is conserved and required for virulence of both species. When introduced into corn cells, Pnss WtsE protein caused water-soaked lesions. In other plants, it either caused cell death or acted as an Avr determinant. Using a yeast- two-hybrid system, WtsE was shown to interact with a number of maize signal transduction proteins that are likely to have roles in either programmed cell death or disease resistance. In Pag and Pab, we have characterized the effector proteins HsvG, HsvB and PthG. HsvG and HsvB are homologous proteins that determine host specificity of Pag and Pab on gypsophila and beet, respectively. Both possess a transcriptional activation domain that functions in yeast. PthG was found to act as an Avr determinant on multiple beet species, but was required for virulence on gypsophila. In addition, we demonstrated that PthG acts within the host cell. Additional effector genes have been characterized on the pathogenicity plasmid, pPATHₚₐg, in Pag. A screen for HrpL- regulated genes in Pnsspointed up 18 candidate effector proteins and four of these were required for full virulence. It is now well established that the virulence of Gram-negative plant pathogenic bacteria is governed by Hrp-dependent effector proteins. However; the mode of action of many effectors is still unresolved. This BARD supported research will significantly contribute to the understanding of how Hrp effectors operate in Pantoea spp. and how they control host specificity and affect symptom production. This may lead to novel approaches for genetically engineering plants resistant to a wide range of bacterial pathogens by inactivating the Hrp effectors with "plantabodies" or modifying their receptors, thereby blocking the induction of the susceptible response. Alternatively, innovative technologies could be used to interfere with the Hrp regulatory cascade by blocking a critical step or mimicking plant or quorum sensing signals.