Journal articles: 'Tral'

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Chu, Chishih, Cheng-Hsun Chiu, Chi-Hong Chu, and Jonathan T. Ou. "Nucleotide and Amino Acid Sequences of oriT-traM-traJ-traY-traA-traL Regions and Mobilization of Virulence Plasmids of Salmonella enterica Serovars Enteritidis, Gallinarum-Pullorum, and Typhimurium." Journal of Bacteriology 184, no. 11 (June 1, 2002): 2857–62. http://dx.doi.org/10.1128/jb.184.11.2857-2862.2002.

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ABSTRACT The virulence plasmid of Salmonella enterica serovar Gallinarum-Pullorum (pSPV) but not those of Salmonella enterica serovars Enteritidis (pSEV) and Typhimurium (pSTV) can be readily mobilized by an F or F-like conjugative plasmid. To investigate the reason for the difference, the oriT-traM-traJ-traY-traA-traL regions of the three salmonella virulence plasmids (pSVs) were cloned and their nucleotide and deduced amino acid sequences were examined. The cloned fragments were generally mobilized more readily than the corresponding full-length pSVs, but the recombinant plasmid containing the oriT of pSPV was, as expected, more readily mobilized, with up to 100-fold higher frequency than the recombinant plasmids containing the oriT of the other two pSVs. The nucleotide sequences of the oriT-traM-traJ-traY-traA-traL region of pSEV and pSTV were almost identical (only 4 bp differences), but differed from that of pSPV. Major nucleotide sequence variations were found in traJ, traY, and the Tra protein binding sites sby and sbm. sby of pSPV showed higher similarity than that of pSEV or pSTV to that of the F plasmid. The reverse was true for sbm: similarity was higher with pSEV and pSTV than with pSPV. In the deduced amino acid sequences of the five Tra proteins, major differences were found in TraY: pSEV's TraY was 75 amino acids, pSTV's was 106 amino acids, and pSPV's was 133 amino acids; and there were duplicate consensus βαα fragments in the TraY of pSPV and F plasmid, whereas there was only a single βαα fragment in that of pSEV and pSTV.

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Karl, Wolfgang, Martina Bamberger, and Ellen L. Zechner. "Transfer Protein TraY of Plasmid R1 Stimulates TraI-Catalyzed oriT Cleavage In Vivo." Journal of Bacteriology 183, no. 3 (February 1, 2001): 909–14. http://dx.doi.org/10.1128/jb.183.3.909-914.2001.

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ABSTRACT The effect of TraY protein on TraI-catalyzed strand scission at the R1 transfer origin (oriT) in vivo was investigated. As expected, the cleavage reaction was not detected in Escherichia coli cells expressing tral and the integration host factor (IHF) in the absence of other transfer proteins. The TraM dependence of strand scission was found to be inversely correlated with the presence of TraY. Thus, the TraY and TraM proteins could each enhance cleaving activity at oriT in the absence of the other. In contrast, no detectable intracellular cleaving activity was exhibited by TraI in an IHF mutant strain despite the additional presence of both TraM and TraY. An essential role for IHF in this reaction in vivo is, therefore, implied. Mobilization experiments employing recombinant R1 oriT constructions and a heterologous conjugative helper plasmid were used to investigate the independent contributions of TraY and TraM to the R1 relaxosome during bacterial conjugation. In accordance with earlier observations,traY was dispensable for mobilization in the presence oftraM, but mobilization did not occur in the absence of bothtraM and traY. Interestingly, although the cleavage assays demonstrate that TraM and TraY independently promote strand scission in vivo, TraM remained essential for mobilization of the R1 origin even in the presence of TraY. These findings suggest that, whereas TraY and TraM function may overlap to a certain extent in the R1 relaxosome, TraM additionally performs a second function that is essential for successful conjugative transmission of plasmid DNA.

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Arutyunov, Denis, Barbara Arenson, Jan Manchak, and Laura S. Frost. "F Plasmid TraF and TraH Are Components of an Outer Membrane Complex Involved in Conjugation." Journal of Bacteriology 192, no. 6 (January 15, 2010): 1730–34. http://dx.doi.org/10.1128/jb.00726-09.

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ABSTRACT F plasmid TraF and TraH are required for F pilus assembly and F plasmid transfer. Using flotation sucrose density gradients, we found that TraF and TraH (as well as TraU and TraW) localized to the outer membrane in the presence of the complete F transfer region, especially TraV, the putative anchor. Mutational analysis of TraH revealed two domains that are important for its function and possible interaction with TrbI, which in turn has a role in stabilizing TraH.

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Tritschler, Felix, Ana Eulalio, Sigrun Helms, Steffen Schmidt, Murray Coles, Oliver Weichenrieder, Elisa Izaurralde, and Vincent Truffault. "Similar Modes of Interaction Enable Trailer Hitch and EDC3 To Associate with DCP1 and Me31B in Distinct Protein Complexes." Molecular and Cellular Biology 28, no. 21 (September 2, 2008): 6695–708. http://dx.doi.org/10.1128/mcb.00759-08.

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ABSTRACT Trailer Hitch (Tral or LSm15) and enhancer of decapping-3 (EDC3 or LSm16) are conserved eukaryotic members of the (L)Sm (Sm and Like-Sm) protein family. They have a similar domain organization, characterized by an N-terminal LSm domain and a central FDF motif; however, in Tral, the FDF motif is flanked by regions rich in charged residues, whereas in EDC3 the FDF motif is followed by a YjeF_N domain. We show that in Drosophila cells, Tral and EDC3 specifically interact with the decapping activator DCP1 and the DEAD-box helicase Me31B. Nevertheless, only Tral associates with the translational repressor CUP, whereas EDC3 associates with the decapping enzyme DCP2. Like EDC3, Tral interacts with DCP1 and localizes to mRNA processing bodies (P bodies) via the LSm domain. This domain remains monomeric in solution and adopts a divergent Sm fold that lacks the characteristic N-terminal α-helix, as determined by nuclear magnetic resonance analyses. Mutational analysis revealed that the structural integrity of the LSm domain is required for Tral both to interact with DCP1 and CUP and to localize to P-bodies. Furthermore, both Tral and EDC3 interact with the C-terminal RecA-like domain of Me31B through their FDF motifs. Together with previous studies, our results show that Tral and EDC3 are structurally related and use a similar mode to associate with common partners in distinct protein complexes.

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Marimo, Patience, Rose Hayeshi, and Stanley Mukanganyama. "Inactivation ofAnopheles gambiaeGlutathione Transferaseε2 by Epiphyllocoumarin." Biochemistry Research International 2016 (2016): 1–8. http://dx.doi.org/10.1155/2016/2516092.

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Glutathione transferases (GSTs) are part of a major family of detoxifying enzymes that can catalyze the reductive dehydrochlorination of dichlorodiphenyltrichloroethane (DDT). The delta and epsilon classes of insect GSTs have been implicated in conferring resistance to this insecticide. In this study, the inactivation ofAnopheles gambiaeGSTε2 by epiphyllocoumarin (Tral 1) was investigated. Recombinant AgGSTε2 was expressed inEscherichia colicells containing a pET3a-AGSTε2 plasmid and purified by affinity chromatography. Tral 1 was shown to inactivate GSTε2 both in a time-dependent manner and in a concentration-dependent manner. The half-life of GSTε2 in the presence of 25 μM ethacrynic acid (ETA) was 22 minutes and with Tral 1 was 30 minutes, indicating that Tral 1 was not as efficient as ETA as an inactivator. The inactivation parameterskinactandKIwere found to be 0.020 ± 0.001 min−1and 7.5 ± 2.1 μM, respectively, after 90 minutes of incubation. Inactivation of GSTε2 by Tral 1 implies that Tral 1 covalently binds to this enzymein vitroand would be expected to exhibit time-dependent effects on the enzymein vivo. Tral 1, therefore, would produce irreversible effects when used together with dichlorodiphenyltrichloroethane (DDT) in malaria control programmes where resistance is mediated by GSTs.

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Ellis, R., and M. Boggild. "Therapy-related acute leukaemia with Mitoxantrone: what is the risk and can we minimise it?" Multiple Sclerosis Journal 15, no. 4 (February 27, 2009): 505–8. http://dx.doi.org/10.1177/1352458508100967.

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Background Therapy-related acute leukaemia (TRAL) is a concern for neurologists and patients when considering treatment with Mitoxantrone for multiple sclerosis (MS). The timing of this complication, risk, mortality and relationship to exposure remain uncertain. Methods We searched literature for publications relating to Mitoxantrone in MS, reviewed publication references and handsearched abstract lists to identify case-series reporting follow-up and complications of treatment with Mitoxantrone. We combined this with our local database of 250 cases treated since 1997. We also identified all reported individual cases of TRAL and extracted data reporting exposure (dose or mg/m2), timing and outcome of TRAL. Results Case-series including 5472 patients were identified; mean dose of Mitoxantrone was 74.2 mg/m2 (range:12–120 mg/m2). TRAL was diagnosed in 0.30% (1 in 333). In 34 TRAL cases, sufficient data was available to inform analysis of exposure. Onset was a median of 18.5 months following Mitoxantrone treatment (range:4–60). Acute Myelocytic Leukaemia and Acute Promyelocytic Leukaemia represented 46.4% each of the leukaemia subtypes. Six of 25 TRAL patients, where outcome was reported, died (24%). Over 80% of cases occurred in patients exposed to >60 mg/m2, with a relative risk of 1.44 (CI95%:1.18–1.70) when comparing total dose >60 mg/m2 against <60 mg/m2 strongly suggesting a relationship between risk of TRAL and total dose.

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Voltz, Raymond, Michaela Starck, Vera Zingler, Michael Strupp, and Hans-Jochem Kolb. "Mitoxantrone therapy in multiple sclerosis and acute leukaemia: a case report out of 644 treated patients." Multiple Sclerosis Journal 10, no. 4 (August 2004): 472–74. http://dx.doi.org/10.1191/1352458504ms1047cr.

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As a rare complication of mitoxantrone (MITOX) therapy in multiple sclerosis (MS), a therapy-related acute leukaemia (TRAL) may develop. The incidence is difficult to estimate, as frequently single cases are reported, up to now a total of eight MS patients. Here we report a new case out of 644 patients. This is a 45-year-old female patient with secondary progressive MS who developed TRAL after a total dose of 48 mg/m2 MITOX. The TRAL was classified as acute myeloblastic leukaemia (AML) M4eo and showed an inversion of chromosome 16 and a partial trisomy 11. Her TRAL was treated with chemotherapy followed by allogeneic bone marrow transplantation. It responded well to the transplantation, whereas the MS symptoms initially worsened but have nearly returned to the pretransplantation level. This report brings the currently published frequency of MITOX-associated TRAL in MS therapy to five in a total of 2336 treated MS patients, representing an incidence of 0.21%.

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Ellis, Richard, Sean Brown, and Mike Boggild. "Therapy-related acute leukaemia with mitoxantrone: Four years on, what is the risk and can it be limited?" Multiple Sclerosis Journal 21, no. 5 (July 10, 2014): 642–45. http://dx.doi.org/10.1177/1352458514541508.

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Therapy-related acute leukaemia (TRAL) is a significant concern, when considering treatment with mitoxantrone for multiple sclerosis (MS). We re-evaluated the literature, identifying all case reports and series of > 50 patients reporting TRAL cases in MS. TRAL was diagnosed in 0.73% of the 12,896 patients identified. Median onset was 22 months following treatment. We calculated a number needed to harm of 137.5 exposed patients, significantly higher than our 2008 analysis. We found that 82.8% of patients were exposed to > 60 mg/m2 with a relative risk of 1.85 ( p = 0.018) compared to < 60mg/m2, strongly suggesting a relationship to dose. MS treatment regimens which limit the mitoxantrone dose to < 60mg/m2 reduce the risk of TRAL.

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Miyazaki, Ryo, Yoshiyuki Ohtsubo, Yuji Nagata, and Masataka Tsuda. "Characterization of the traD Operon of Naphthalene-Catabolic Plasmid NAH7: a Host-Range Modifier in Conjugative Transfer." Journal of Bacteriology 190, no. 19 (August 1, 2008): 6281–89. http://dx.doi.org/10.1128/jb.00709-08.

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ABSTRACT Pseudomonas putida G7 carries a naphthalene-catabolic and self-transmissible plasmid, NAH7, which belongs to the IncP-9 incompatibility group. Adjacent to the putative origin of conjugative transfer (oriT) of NAH7 are three genes, traD, traE, and traF, whose functions and roles in conjugation were previously unclear. These three genes were transcribed monocistronically and thus were designated the traD operon. Mutation of the three genes in the traD operon resulted in 10- to 105-fold decreases in the transfer frequencies of the plasmids from Pseudomonas to Pseudomonas and Escherichia coli and from E. coli to E. coli. On the other hand, the traD operon was essential for the transfer of NAH7 from E. coli to Pseudomonas strains. These results indicated that the traD operon is a host-range modifier in the conjugative transfer of NAH7. The TraD, TraE, and TraF proteins were localized in the cytoplasm, periplasm, and membrane, respectively, in strain G7 cells. Our use of a bacterial two-hybrid assay system showed that TraE interacted in vivo with other essential components for conjugative transfer, including TraB (coupling protein), TraC (relaxase), and MpfH (a channel subunit in the mating pair formation system).

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Schaper, Elke, Alexander Korsunsky, Jūlija Pečerska, Antonio Messina, Riccardo Murri, Heinz Stockinger, Stefan Zoller, Ioannis Xenarios, and Maria Anisimova. "TRAL: tandem repeat annotation library." Bioinformatics 31, no. 18 (May 18, 2015): 3051–53. http://dx.doi.org/10.1093/bioinformatics/btv306.

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11

Childers, Donald G., Ira S. Fischler, Timothy L. Boaz, Nathan W. Perry, and A. Antonio Arroyo. "Multichannel, Single Tral Event Related Potential Classification." IEEE Transactions on Biomedical Engineering BME-33, no. 12 (December 1986): 1069–75. http://dx.doi.org/10.1109/tbme.1986.325683.

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12

Harris, Robin L., and Philip M. Silverman. "Tra Proteins Characteristic of F-Like Type IV Secretion Systems Constitute an Interaction Group by Yeast Two-Hybrid Analysis." Journal of Bacteriology 186, no. 16 (August 15, 2004): 5480–85. http://dx.doi.org/10.1128/jb.186.16.5480-5485.2004.

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ABSTRACT Using yeast two-hybrid screens, we have defined an interaction group of six Tra proteins encoded by the F plasmid and required by F+ cells to elaborate F pili. The six proteins are TraH, TraF, TraW, TraU, TrbI, and TrbB. Except for TrbI, these proteins were all identified as hallmarks of F-like type IV secretion systems (TFSSs), with no homologues among TFSS genes of P-type or I-type systems (T. Lawley, W. Klimke, M. Gubbins, and L. Frost, FEMS Microbiol. Lett. 224:1-15, 2003). Also with the exception of TrbI, which is an inner membrane protein, the remaining proteins are or are predicted to be periplasmic. TrbI consists of one membrane-spanning segment near its N terminus and an 88-residue, hydrophilic domain that extends into the periplasm. Hence, the proteins of this group probably form a periplasmic cluster in Escherichia coli. The interaction network identifies TraH as the most highly connected node, with two-hybrid links to TrbI, TraU, and TraF. As measured by transcriptional activation of lacZ, the TrbI-TraH interaction in Saccharomyces cerevisiae requires the TraH amino acid segment from residues 193 to 225. The TraU and TraF interactions are localized to C-terminal segments of TraH (amino acids 315 to 458 for TraF and amino acids 341 to 458 for TraU). The TrbI-TraH interaction with full-length (less the signal peptide) TraH is weak but increases 40-fold with N-terminal TraH deletions; the first 50 amino acids appear to be critical for inhibiting TrbI binding in yeast. Previous studies by others have shown that, with the exception of trbB mutations, which do not affect the elaboration or function of F pili under laboratory conditions, a mutation in any of the other genes in this interaction group alters the number or length distribution of F pili. We propose a model whereby one function of the TraH interaction group is to control F-pilus extension and retraction.

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Aijmer, Göran. "Rice, Death, and Chiefly Power in Cen- tral Borneo." Anthropos 105, no. 2 (2010): 393–410. http://dx.doi.org/10.5771/0257-9774-2010-2-393.

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14

Grahn, A. Marika, Jana Haase, Dennis H. Bamford, and Erich Lanka. "Components of the RP4 Conjugative Transfer Apparatus Form an Envelope Structure Bridging Inner and Outer Membranes of Donor Cells: Implications for Related Macromolecule Transport Systems." Journal of Bacteriology 182, no. 6 (March 15, 2000): 1564–74. http://dx.doi.org/10.1128/jb.182.6.1564-1574.2000.

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ABSTRACT During bacterial conjugation, the single-stranded DNA molecule is transferred through the cell envelopes of the donor and the recipient cell. A membrane-spanning transfer apparatus encoded by conjugative plasmids has been proposed to facilitate protein and DNA transport. For the IncPα plasmid RP4, a thorough sequence analysis of the gene products of the transfer regions Tra1 and Tra2 revealed typical features of mainly inner membrane proteins. We localized essential RP4 transfer functions to Escherichia coli cell fractions by immunological detection with specific polyclonal antisera. Each of the gene products of the RP4 mating pair formation (Mpf) system, specified by the Tra2 core region and by traF of the Tra1 region, was found in the outer membrane fraction with one exception, the TrbB protein, which behaved like a soluble protein. The membrane preparation from Mpf-containing cells had an additional membrane fraction whose density was intermediate between those of the cytoplasmic and outer membranes, suggesting the presence of attachment zones between the twoE. coli membranes. The Tra1 region is known to encode the components of the RP4 relaxosome. Several gene products of this transfer region, including the relaxase TraI, were detected in the soluble fraction, but also in the inner membrane fraction. This indicates that the nucleoprotein complex is associated with and/or assembled facing the cytoplasmic site of the E. coli cell envelope. The Tra1 protein TraG was predominantly localized to the cytoplasmic membrane, supporting its potential role as an interface between the RP4 Mpf system and the relaxosome.

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Schmidt-Eisenlohr, Heike, Natalie Domke, and Christian Baron. "TraC of IncN Plasmid pKM101 Associates with Membranes and Extracellular High-Molecular-Weight Structures inEscherichia coli." Journal of Bacteriology 181, no. 18 (September 15, 1999): 5563–71. http://dx.doi.org/10.1128/jb.181.18.5563-5571.1999.

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ABSTRACT Conjugative transfer of IncN plasmid pKM101 is mediated by the TraI-TraII region-encoded transfer machinery components. Similar to the case for the related Agrobacterium tumefaciens T-complex transfer apparatus, this machinery is needed for assembly of pili to initiate cell-to-cell contact preceding DNA transfer. Biochemical and cell biological experiments presented here show extracellular localization of TraC, as suggested by extracellular complementation of TraC-deficient bacteria by helper cells expressing a functional plasmid transfer machinery (S. C. Winans, and G. C. Walker, J. Bacteriol. 161:402–410, 1985). Overexpression of TraC and its export in large amounts into the periplasm of Escherichia coliallowed purification by periplasmic extraction, ammonium sulfate precipitation, and column chromatography. Whereas TraC was soluble in overexpressing strains, it partly associated with the membranes in pKM101-carrying cells, possibly due to protein-protein interactions with other components of the TraI-TraII region-encoded transfer machinery. Membrane association of TraC was reduced in strains carrying pKM101 derivatives with transposon insertions in genes coding for other essential components of the transfer machinery,traM, traB, traD, andtraE but not eex, coding for an entry exclusion protein not required for DNA transfer. Cross-linking identified protein-protein interactions of TraC in E. coli carrying pKM101 but not derivatives with transposon insertions in essentialtra genes. Interactions with membrane-bound Tra proteins may incorporate TraC into a surface structure, suggested by its removal from the cell by shearing as part of a high-molecular-weight complex. Heterologous expression of TraC in A. tumefaciens partly compensated for the pilus assembly defect in strains deficient for its homolog VirB5, which further supported its role in assembly of conjugative pili. In addition to its association with high-molecular-weight structures, TraC was secreted into the extracellular milieu. Conjugation experiments showed that secreted TraC does not compensate transfer deficiency of TraC-deficient cells, suggesting that extracellular complementation may rely on cell-to-cell transfer of TraC only as part of a bona fide transfer apparatus.

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Lawley, Trevor D., Matthew W. Gilmour, James E. Gunton, Leah J. Standeven, and Diane E. Taylor. "Functional and Mutational Analysis of Conjugative Transfer Region 1 (Tra1) from the IncHI1 Plasmid R27." Journal of Bacteriology 184, no. 8 (April 15, 2002): 2173–80. http://dx.doi.org/10.1128/jb.184.8.2173-2180.2002.

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ABSTRACT The conjugative transfer region 1 (Tra1) of the IncHI1 plasmid R27 was subjected to DNA sequence analysis, mutagenesis, genetic complementation, and an H-pilus-specific phage assay. Analysis of the nucleotide sequence indicated that the Tra1 region contains genes coding for mating pair formation (Mpf) and DNA transfer replication (Dtr) and a coupling protein. Insertional disruptions of 9 of the 14 open reading frames (ORFs) in the Tra1 region resulted in a transfer-deficient phenotype. Conjugative transfer was restored for each transfer mutant by genetic complementation. An intergenic region between traH and trhR was cloned and mobilized by R27, indicating the presence of an origin of transfer (oriT). The five ORFs immediately downstream of the oriT region are involved in H-pilus production, as determined by an H-pilus-specific phage assay. Three of these ORFs encode proteins homologous to Mpf proteins from IncF plasmids. Upstream of the oriT region are four ORFs required for plasmid transfer but not H-pilus production. TraI contains sequence motifs that are characteristic of relaxases from the IncP lineage but share no overall homology to known relaxases. TraJ contains both an Arc repressor motif and a leucine zipper motif. A putative coupling protein, TraG, shares a low level of homology to the TraG family of coupling proteins and contains motifs that are important for DNA transfer. This analysis indicates that the Mpf components of R27 share a common lineage with those of the IncF transfer system, whereas the relaxase of R27 is ancestrally related to that of the IncP transfer system.

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Pelechov�, Jitka. "Le paysage th��tral berlinois depuis les ann�es 1990." �tudes th��trales N�58, no. 3 (2013): 25. http://dx.doi.org/10.3917/etth.058.0025.

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Tkachyck, Pavlo, and Alexandr Yemelianov. "Influence of soil characteristics on working dynamics mining tral authority." Military Technical Collection, no. 24 (May 20, 2021): 46–51. http://dx.doi.org/10.33577/2312-4458.24.2021.46-51.

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The safest way to clear mines and barriers, in terms of saving human life and health, is to clear them at the site. Minesweepers are being used for this purpose, and in recent years they have gained extensive research, which is aimed at the use of large-scale explosion technology. Existing minesweepers in the Armed Forces of Ukraine, as a rule, are old-fashioned, and have certain shortcomings that affect the efficiency of their direct use. These include, first of all, insufficient survivability and different efficiency of their use during demining of minefields with different types of soils: from sandy, loamy and gravel. The range of physical and mechanical properties of the latter is very wide, and therefore - for the operation of an explosive device (mine) requires different amounts of action on the soil surface. The modernized mine trawl neutralizing device in the form of a system of U-shaped rockers with disks at the end is proposed in the work. They, acting directly on the mine, or through a small layer of soil, cause it to explode, provided that the force on the latter is not less than the minimum value required for its disposal. The advantage of this type of mine trawl is that even if one of the working disks fails during the explosion, it is structurally relatively easy to replace it with another. The magnitude of the deepening of the neutralizing disks into the soil (at a constant weight of the mounted part of the trawl), as well as their dynamic action through the soil on the mine depends on the physical and mechanical properties of the soil. Therefore, for the case of, for example, clay or sandy soils during the movement of the trawl, it will be sufficient for the operation of the explosive device, at the same time for the coating of gravel - insufficient. In addition, the amount of deepening of the working disks in the soil depends on its humidity. The study of the influence of the main physical and mechanical characteristics of soils on the dynamics of the modernized mine trawl neutralizing device during movement along the minefield (before and after the mine explosion) is the subject of research, hence their relevance.

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Kuryshova, Yulia. "An inverse spectral problem for differential operators with integral delay." Tamkang Journal of Mathematics 42, no. 3 (August 24, 2011): 295–303. http://dx.doi.org/10.5556/j.tkjm.42.2011.743.

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The uniqueness theorem is proved for the solution of the inverse spec- tral problem for second-order integro-di®erential operators on a ¯nite interval. These operators are perturbations of the Sturm-Liouville operator with convolution and one- dimensional operators. The main tool is an integral transform connected with solutions of integro-di®erential operators.

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Lin, Mei-Hui, and Shih-Tung Liu. "Stabilization of pSW100 from Pantoea stewartii by the F Conjugation System." Journal of Bacteriology 190, no. 10 (March 14, 2008): 3681–89. http://dx.doi.org/10.1128/jb.00846-07.

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ABSTRACT Plasmid pSW100 is 1 of the 13 plasmids from Pantoea stewartii subsp. stewartii SW2 which has a replicon that resembles that of ColE1. This work uses a pSW100 derivative, pSW140K, to study how the pSW100 replicon is stably maintained in its hosts. Our results indicate that although pSW140K is stable in Escherichia coli HB101, the plasmid is rapidly lost in another E. coli strain, DH5α, indicating that the genetic background of an E. coli strain affects the stability of pSW140K. Mutagenesis of E. coli HB101 with EZ::TN <DHFR-1> revealed that mutations in traC, traF, traG, traN, and traV, which encode the components of the sex pilus assembly, reduce plasmid stability. Furthermore, this work identified that a 38-bp region located immediately upstream of the RNAII promoter is critical to the maintenance of plasmid stability in E. coli HB101. TraC binds to the region, and in addition, deleting the region destabilizes the plasmid. Furthermore, inserting this 38-bp fragment into a plasmid that contains the minimal replicon from pSW200 stabilizes the plasmid in E. coli HB101. Fluorescence in situ hybridization and immunofluorescence staining also revealed that derivatives of pSW100, pSW128A, and TraC are colocalized in cells, suggesting that pSW100 may use the sex pilus assembly as a partition apparatus to ensure the even distribution of the plasmid during cell division, which may thus maintain the plasmid's stability.

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He, Xuesong, William Chang, Deanne L. Pierce, Laura Ort Seib, Jennifer Wagner, and Clay Fuqua. "Quorum Sensing in Rhizobium sp. Strain NGR234 Regulates Conjugal Transfer (tra) Gene Expression and Influences Growth Rate." Journal of Bacteriology 185, no. 3 (February 1, 2003): 809–22. http://dx.doi.org/10.1128/jb.185.3.809-822.2003.

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ABSTRACT Rhizobium sp. strain NGR234 forms symbiotic, nitrogen-fixing nodules on a wide range of legumes via functions largely encoded by the plasmid pNGR234a. The pNGR234a sequence revealed a region encoding plasmid replication (rep) and conjugal transfer (tra) functions similar to those encoded by the rep and tra genes from the tumor-inducing (Ti) plasmids of Agrobacterium tumefaciens, including homologues of the Ti plasmid quorum-sensing regulators TraI, TraR, and TraM. In A. tumefaciens, TraI, a LuxI-type protein, catalyzes synthesis of the acylated homoserine lactone (acyl-HSL) N-3-oxo-octanoyl-l-homoserine lactone (3-oxo-C8-HSL). TraR binds 3-oxo-C8-HSL and activates expression of Ti plasmid tra and rep genes, increasing conjugation and copy number at high population densities. TraM prevents this activation under noninducing conditions. Although the pNGR234a TraR, TraI, and TraM appear to function similarly to their A. tumefaciens counterparts, the TraR and TraM orthologues are not cross-functional, and the quorum-sensing systems have differences. NGR234 TraI synthesizes an acyl-HSL likely to be 3-oxo-C8-HSL, but traI mutants and a pNGR234a-cured derivative produce low levels of a similar acyl-HSL and another, more hydrophobic signal molecule. TraR activates expression of several pNGR234a tra operons in response to 3-oxo-C8-HSL and is inhibited by TraM. However, one of the pNGR234a tra operons is not activated by TraR, and conjugal efficiency is not affected by TraR and 3-oxo-C8-HSL. The growth rate of NGR234 is significantly decreased by TraR and 3-oxo-C8-HSL through functions encoded elsewhere in the NGR234 genome.

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Martucci, Giovanni, Francisco Navas-Guzmán, Ludovic Renaud, Gonzague Romanens, S. Mahagammulla Gamage, Maxime Hervo, Pierre Jeannet, and Alexander Haefele. "Validation of pure rotational Raman temperature data from the Raman Lidar for Meteorological Observations (RALMO) at Payerne." Atmospheric Measurement Techniques 14, no. 2 (February 22, 2021): 1333–53. http://dx.doi.org/10.5194/amt-14-1333-2021.

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Abstract. The Raman Lidar for Meteorological Observations (RALMO) is operated at the MeteoSwiss station of Payerne (Switzerland) and provides, amongst other products, continuous measurements of temperature since 2010. The temperature profiles are retrieved from the pure rotational Raman (PRR) signals detected around the 355 nm Cabannes line. The transmitter and receiver systems of RALMO are described in detail, and the reception and acquisition units of the PRR channels are thoroughly characterized. The FastCom P7888 card used to acquire the PRR signal, the calculation of the dead time and the desaturation procedure are also presented. The temperature profiles retrieved from RALMO PRR data during the period going from July 2017 to the end of December 2018 have been validated against two reference operational radiosounding systems (ORSs) co-located with RALMO, i.e. the Meteolabor SRS-C50 and the Vaisala RS41. The ORSs have also served to perform the calibration of the RALMO temperature during the validation period. The maximum bias (ΔTmax), mean bias (μ) and mean standard deviation (σ) of RALMO temperature Tral with respect to the reference ORS, Tors, are used to characterize the accuracy and precision of Tral along the troposphere. The daytime statistics provide information essentially about the lower troposphere due to lower signal-to-noise ratio. The ΔTmax, μ and σ of the differences ΔT=Tral-Tors are, respectively, 0.28, 0.02±0.1 and 0.62±0.03 K. The nighttime statistics provide information for the entire troposphere and yield ΔTmax=0.29 K, μ=0.05±0.34 K and σ=0.66±0.06 K. The small ΔTmax, μ and σ values obtained for both daytime and nighttime comparisons indicate the high stability of RALMO that has been calibrated only seven times over 18 months. The retrieval method can correct for the largest sources of correlated and uncorrelated errors, e.g. signal noise, dead time of the acquisition system and solar background. Especially the solar radiation (scattered into the field of view from the zenith angle Φ) affects the quality of PRR signals and represents a source of systematic error for the retrieved temperature. An imperfect subtraction of the background from the daytime PRR profiles induces a bias of up to 2 K at all heights. An empirical correction f(Φ) ranging from 0.99 to 1 has therefore been applied to the mean background of the PRR signals to remove the bias. The correction function f(Φ) has been validated against the numerical weather prediction model COSMO (Consortium for Small-scale Modelling), suggesting that f(Φ) does not introduce any additional source of systematic or random error to Tral. A seasonality study has been performed to help with understanding if the overall daytime and nighttime zero bias hides seasonal non-zero biases that cancel out when combined in the full dataset.

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Shakeyev, Kairat. "Theoretical basing of endoprosthesis replacement for paracolostomal and ven-tral hernias." Perspectives of Innovations, Economics and Business 2, no. 2 (October 9, 2009): 149–52. http://dx.doi.org/10.15208/pieb.2009.86.

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Harari, Josu�. "Artaudet le th��tre Balinais: imaginaire th��tral ou fait ethnologique." Neophilologus 71, no. 1 (January 1987): 55–65. http://dx.doi.org/10.1007/bf00556704.

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Fekete, Richard A., and Laura S. Frost. "Mobilization of Chimeric oriT Plasmids by F and R100-1: Role of Relaxosome Formation in Defining Plasmid Specificity." Journal of Bacteriology 182, no. 14 (July 15, 2000): 4022–27. http://dx.doi.org/10.1128/jb.182.14.4022-4027.2000.

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ABSTRACT Cleavage at the F plasmid nic site within the origin of transfer (oriT) requires the F-encoded proteins TraY and TraI and the host-encoded protein integration host factor in vitro. We confirm that F TraY, but not F TraM, is required for cleavage atnic in vivo. Chimeric plasmids were constructed which contained either the entire F or R100-1 oriT regions or various combinations of nic, TraY, and TraM binding sites, in addition to the traM gene. The efficiency of cleavage atnic and the frequency of mobilization were assayed in the presence of F or R100-1 plasmids. The ability of these chimeric plasmids to complement an F traM mutant or affect F transfer via negative dominance was also measured using transfer efficiency assays. In cases where cleavage at nic was detected, R100-1 TraI was not sensitive to the two-base difference in sequence immediately downstream of nic, while F TraI was specific for the F sequence. Plasmid transfer was detected only when TraM was able to bind to its cognate sites within oriT. High-affinity binding of TraY in cis to oriTallowed detection of cleavage at nic but was not required for efficient mobilization. Taken together, our results suggest that stable relaxosomes, consisting of TraI, -M, and -Y bound to oriT are preferentially targeted to the transfer apparatus (transferosome).

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Popelard, Marie-Dominique. "Remarques sur le m�tissage des voix dans le dialoguer th��tral." �tudes th��trales N�31-32, no. 2 (2004): 114. http://dx.doi.org/10.3917/etth.031.0114.

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Chiappone-Lucchesi, Magali. "Le t�moignage th��tral de Charlotte Delbo�: du double au testament." �tudes th��trales N�51-52, no. 2 (2011): 33. http://dx.doi.org/10.3917/etth.051.0033.

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Roosen, Tanguy. "La gestion collective du droit d�auteur dans le domaine th��tral." �tudes th��trales N�62, no. 1 (2015): 43. http://dx.doi.org/10.3917/etth.062.0043.

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Höppner, Christoph, Zhenying Liu, Natalie Domke, Andrew N. Binns, and Christian Baron. "VirB1 Orthologs from Brucella suis and pKM101 Complement Defects of the Lytic Transglycosylase Required for Efficient Type IV Secretion from Agrobacterium tumefaciens." Journal of Bacteriology 186, no. 5 (March 1, 2004): 1415–22. http://dx.doi.org/10.1128/jb.186.5.1415-1422.2004.

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ABSTRACT Type IV secretion systems mediate conjugative plasmid transfer as well as the translocation of virulence factors from various gram-negative pathogens to eukaryotic host cells. The translocation apparatus consists of 9 to 12 components, and the components from different organisms are believed to have similar functions. However, orthologs to proteins of the prototypical type IV system, VirB of Agrobacterium tumefaciens, typically share only 15 to 30% identical amino acids, and functional complementation between components of different type IV secretion systems has not been achieved. We here report a heterologous complementation in the case of A. tumefaciens virB1 defects with its orthologs from Brucella suis (VirB1s) and the IncN plasmid pKM101 (TraL). In contrast, expression of the genes encoding the VirB1 orthologs from the IncF plasmid (open reading frame 169) and from the Helicobacter pylori cag pathogenicity island (HP0523) did not complement VirB1 functions. The complementation of VirB1 activity was assessed by T-pilus formation, by tumor formation on wounded plants, by IncQ plasmid transfer, and by IncQ plasmid recipient assay. Replacement of the key active-site Glu residue by Ala abolished the complementation by VirB1 from B. suis and by TraL, demonstrating that heterologous complementation requires an intact lytic transglycosylase active site. In contrast, the VirB1 active-site mutant from A. tumefaciens retained considerable residual activity in various activity assays, implying that this protein exerts additional effects during the type IV secretion process.

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Teng, C. T., C. de la Fuente-Sandoval, C. Lupo, R. Córdoba, P. Cabrera, A. Pfau, and B. Soares. "PMH10 TREATMENT-RESISTANT DEPRESSION IN LATIN AMERICA: PRELIMINARY RESULTS FROM THE TRAL STUDY." Value in Health Regional Issues 19 (October 2019): S50—S51. http://dx.doi.org/10.1016/j.vhri.2019.08.291.

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Moldofsky, Harvey. "The Significance, Assessment, and Management of Nonrestorative Sleep in Fibromyalgia Syndrome." CNS Spectrums 13, S5 (March 2008): 22–26. http://dx.doi.org/10.1017/s1092852900026808.

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AbstractPeople with fibromyalgia syndrome (FMS) experi-ence unrefreshing sleep, aches, hypersensitivity, and cognitive and emotional difficulties. Although no specific causative factor or biological agent is known to account for all of the features of FMS and these related diagnoses, the generalized hypersensitivity of the body is considered to be affected by disturbances in cen-tral nervous system (CNS) functions. Such CNS dis-turbances are intrinsic to the sleeping-waking brain, where the common symptom elements in all these illnesses are poor quality of sleep, nonspecific pain, fatigue, and psychological distress in the absence of known disease pathology.

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Gala, Candelas. "La estética cuántica:Juan Larrea, Niels Bohr y el tercero incluido." Çédille, no. 18 (2020): 203–33. http://dx.doi.org/10.25145/j.cedille.2020.18.09.

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This essay explores notable coincidences in Juan Larrea’s poetics with some cen-tral points in quantum physics, such as, the correlation between the search in his inner self and the investigations on the atom taking place during the first decades of the twentieth-century, between leaving aside the monolithic subjectivity in favor of a universal and collective Spirit and the «entangled» observer in the cosmic web, object of his/her obser-vation, in modern physics, and, particularly, the role of the French language in dealing with the conflict with Western dualisms and their possible resolution in Bohr’s principle of complementarity, a parallel of Basarab Nicolescu’s theory of the hidden third.

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Will, William R., and Laura S. Frost. "Hfq Is a Regulator of F-Plasmid TraJ and TraM Synthesis in Escherichia coli." Journal of Bacteriology 188, no. 1 (January 1, 2006): 124–31. http://dx.doi.org/10.1128/jb.188.1.124-131.2006.

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ABSTRACT The F plasmid of Escherichia coli allows horizontal DNA transfer between an F+ donor cell and an F− recipient. Expression of the transfer genes is tightly controlled by a number of factors, including the following plasmid-encoded regulatory proteins: TraJ, the primary activator of the 33-kb tra operon, and the autoregulators TraM and TraY. Here, we demonstrate that the host RNA binding protein, Hfq, represses TraJ and TraM synthesis by destabilizing their respective mRNAs. Mating assays and immunoblot analyses for TraM and TraJ showed that transfer efficiency and protein levels increased in host cells containing a disruption in hfq compared to wild-type cells in stationary phase. The stability of transcripts containing a putative Hfq binding site located in the intergenic untranslated region between traM and traJ was increased in hfq mutant donor cells, suggesting that Hfq destabilizes these transcripts. Electrophoretic mobility shift assays demonstrated that Hfq specifically binds this region but not the antisense RNA, FinP, encoded on the opposite strand. Together, these findings indicate that Hfq regulates traM and traJ transcript stability by a mechanism separate from FinOP-mediated repression.

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Sanabria, Marta, Laura Morel, and Gloria Aguilar. "General Characteristics of Lactation Centers in Reference Hospitals of Asunción and the Cen-tral Department." Pediatría (Asunción) 44, no. 2 (August 30, 2017): 143–47. http://dx.doi.org/10.18004/ped.2017.agosto.143-147.

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Zhang, J., Ik-Hwan Kim, S. J. Hong, and D. S. Seo. "High-speed optical pulse train generation based on line-by-line spec-tral intensity coding." Ukrainian Journal of Physical Optics 12, no. 3 (2011): 117. http://dx.doi.org/10.3116/16091833/12/3/117/2011.

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Knútsdóttir, Vera. "SPECTRAL MEMORIES: AESTHETIC RESPONSES TO THE FINANCIAL CRASH IN ICELAND 2008." Nordic Journal of Aesthetics 29, no. 60 (November 22, 2020): 116–39. http://dx.doi.org/10.7146/nja.v29i60.122844.

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In October 2008, one of the largest bank crashes in history struck Iceland, a country of three hundred and thirty five thousand inhab-itants. The aim of the article is to examine two cultural responses to the crash and the crisis that followed. More precisely, the aim is to analyse how the creation of the haunted house in I Remember You, a crash-horror story by crime writer Yrsa Sigurðardóttir, as well as the spectral half-built houses portrayed by visual artist Guðjón Ketilsson refer quite directly, yet spectrally, to the period. The spec-tral themes of the two works give the opportunity to discuss the moment following the crash as a moment of haunting—but who is haunted and by whom?

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Schröder, Gunnar, Sabine Krause, Ellen L. Zechner, Beth Traxler, Hye-Jeong Yeo, Rudi Lurz, Gabriel Waksman, and Erich Lanka. "TraG-Like Proteins of DNA Transfer Systems and of the Helicobacter pylori Type IV Secretion System: Inner Membrane Gate for Exported Substrates?" Journal of Bacteriology 184, no. 10 (May 15, 2002): 2767–79. http://dx.doi.org/10.1128/jb.184.10.2767-2779.2002.

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ABSTRACT TraG-like proteins are potential NTP hydrolases (NTPases) that are essential for DNA transfer in bacterial conjugation. They are thought to mediate interactions between the DNA-processing (Dtr) and the mating pair formation (Mpf) systems. TraG-like proteins also function as essential components of type IV secretion systems of several bacterial pathogens such as Helicobacter pylori. Here we present the biochemical characterization of three members of the family of TraG-like proteins, TraG (RP4), TraD (F), and HP0524 (H. pylori). These proteins were found to have a pronounced tendency to form oligomers and were shown to bind DNA without sequence specificity. Standard NTPase assays indicated that these TraG-like proteins do not possess postulated NTP-hydrolyzing activity. Surface plasmon resonance was used to demonstrate an interaction between TraG and relaxase TraI of RP4. Topology analysis of TraG revealed that TraG is a transmembrane protein with cytosolic N and C termini and a short periplasmic domain close to the N terminus. We predict that multimeric inner membrane protein TraG forms a pore. A model suggesting that the relaxosome binds to the TraG pore via TraG-DNA and TraG-TraI interactions is presented.

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PANG, Zhiwei, Binduo XU, Xuegang CHEN, and Yiping REN. "Community structure and diversity of shrimp assemblages in the cen-tral waters of Jiaozhou Bay, China." Journal of Fishery Sciences of China 20, no. 2 (December 16, 2013): 361–71. http://dx.doi.org/10.3724/sp.j.1118.2013.00361.

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Yang, Zhi-xian, Xiang-wei Yu, Yue-jun Zheng, Yun-tai Chen, Xiao-xi Ni, and Winston Chan. "Earthquake relocation and 3-dimensional crustal structure of P-wave velocity in cen-tral-western China." Acta Seismologica Sinica 17, no. 1 (January 2004): 20–30. http://dx.doi.org/10.1007/bf03191391.

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Tun-Garrido, Cristina, Patricia Bustos, Víctor González, and Susana Brom. "Conjugative Transfer of p42a from Rhizobium etli CFN42, Which Is Required for Mobilization of the Symbiotic Plasmid, Is Regulated by Quorum Sensing." Journal of Bacteriology 185, no. 5 (March 1, 2003): 1681–92. http://dx.doi.org/10.1128/jb.185.5.1681-1692.2003.

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ABSTRACT Rhizobium etli CFN42 contains six plasmids. Only one of them, p42a, is self-conjugative at high frequency. This plasmid is strictly required for mobilization of the symbiotic plasmid (pSym). To study the transfer mechanism of p42a, a self-transmissible cosmid clone containing its transfer region was isolated. Its sequence showed that most of the tra genes are highly similar to genes of Agrobacterium tumefaciens pTiC58 and other related plasmids. Four putative regulatory genes were identified; three of these (traI, traR, and cinR) belong to the LuxR-LuxI family. Mutagenesis of these genes confirmed their requirement for p42a transfer. We found that the conjugative transfer of p42a is dependent on quorum sensing, and consequently pSym transfer also was found to be similarly regulated, establishing a complex link between environmental conditions and pSym transfer. Although R. etli has been shown to produce different N-acyl-homoserine lactones, only one of them, a 3-oxo-C8-homoserine lactone encoded by the traI gene described here, was involved in transfer. Mutagenesis of the fourth regulatory gene, traM, had no effect on transfer. Analysis of transcriptional fusions of the regulatory genes to a reporter gene suggests a complex regulation scheme for p42a conjugative transfer. Conjugal transfer gene expression was found to be directly upregulated by TraR and the 3-oxo-C8-homoserine lactone synthesized by TraI. The traI gene was autoregulated by these elements and positively regulated by CinR, while cinR expression required traI. Finally, we did not detect expression of traM, indicating that in p42a TraM may be expressed so weakly that it cannot inhibit conjugal transfer, leading to the unrepressed transfer of p42a.

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García Sudo, Alejandro. "Cuando la Banda de la Marina Estadounidense tocaba al compás panamericano: esbozo de los albores del intercambio musical en el sistema interamericano (1924-1933)." Revista de Historia de América, no. 156 (January 30, 2019): 351–70. http://dx.doi.org/10.35424/rha.156.2019.249.

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Una versión preliminar del documento que se presenta a continuación seleyó como ponencia el 30 de abril de 2017 en la reunión anual de la Latin American Studies Association (LASA) como parte de la mesa titulada “Re-pensar el panamericanismo de la primera mitad del siglo XX”. La idea cen-tral es que el análisis de la programación y la ejecución de la música que se tocó en actos oficiales de la Unión Panamericana ayudan a concebir el mo-vimiento panamericano como un conjunto de prácticas socioculturales en la esfera pública internacional. El autor comparte el texto como un avance deinvestigación para dar a conocer algunas de las líneas temáticas de su tesisdoctoral en musicología, que defenderá próximamente en la Universidad deCalifornia, Los Ángeles (UCLA).

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Paterson, E. Suzanne, Margret I. Moré, Gansen Pillay, Christina Cellini, Roger Woodgate, Graham C. Walker, V. N. Iyer, and Stephen C. Winans. "Genetic Analysis of the Mobilization and Leading Regions of the IncN plasmids pKM101 and pCU1." Journal of Bacteriology 181, no. 8 (April 15, 1999): 2572–83. http://dx.doi.org/10.1128/jb.181.8.2572-2583.1999.

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ABSTRACT The conjugative IncN plasmids pKM101 and pCU1 have previously been shown to contain identical oriT sequences as well as conserved restriction endonuclease cleavage patterns within theirtra regions. Complementation analysis and sequence data presented here indicate that these two plasmids encode essentially identical conjugal DNA-processing proteins. This region contains three genes, traI, traJ, and traK, transcribed in the same orientation from a promoter that probably lies within or near the conjugal transfer origin (oriT). Three corresponding proteins were visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and complementation analysis confirmed that this region contains three tracomplementation groups. All three proteins resemble proteins of the IncW plasmid R388 and other plasmids thought to have roles in processing of plasmid DNA during conjugation. The hydropathy profile of TraJ suggests a transmembrane topology similar to that of several homologous proteins. Both traK and traI were required for efficient interplasmid site-specific recombination atoriT, while traJ was not required. The leading region of pKM101 contains three genes (stbA,stbB, and stbC), null mutations in which cause elevated levels of plasmid instability. Plasmid instability was observed only in hosts that are proficient in interplasmid recombination, suggesting that this recombination can potentially lead to plasmid loss and that Stb proteins somehow overcome this, possibly via site-specific multimer resolution.

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Lubik-Reczek, Natasza. "Transformation of the Countries of Cen- tral and Eastern Europe – an Attempt at Comparing Croatia and Slovenia." Przegląd Politologiczny, no. 3 (September 1, 2015): 79. http://dx.doi.org/10.14746/pp.2015.20.3.6.

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Smith, C. Jeffrey, and Anita C. Parker. "A gene product related to Tral is required for the mobilization of Bacteroides mobilizable transposons and plasmids." Molecular Microbiology 20, no. 4 (May 1996): 741–50. http://dx.doi.org/10.1111/j.1365-2958.1996.tb02513.x.

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Inamoto, Susumu, Hirokazu Fukuda, Tatsuhiko Abo, and Eiichi Ohtsubo. "Site- and Strand-Specific Nicking at oriT of Plasmid R100 in a Purified System: Enhancement of the Nicking Activity of Tral (Helicase I) with TraY and IHF1." Journal of Biochemistry 116, no. 4 (October 1994): 838–44. http://dx.doi.org/10.1093/oxfordjournals.jbchem.a124604.

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46

Asmael, M. B. A., Roslee Ahmad, Ali Ourdjini, and S. Farahany. "Effect of Elements Cerium and Lanthanum on Eutectic Solidification of Al-Si-Cu near Eutectic Cast Alloy." Advanced Materials Research 845 (December 2013): 118–22. http://dx.doi.org/10.4028/www.scientific.net/amr.845.118.

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The properties of Al-Si-Cu cast alloys are strongly affected by eutectic Al-Si and Al-Cu phases. The characteristic parameters of these two phases with additions cerium 1wt % (Ce) and lanthanum1 wt % (La) were investigated in Al-11Si-2Cu near eutectic alloy using computer-aided cooling curve thermal analysis. As a result, the La additive showed the highest (TNAl-Si) while the Ce additive showed very little effect. In addition, the growth temperature (TGAl-Si) is decreased by adding Ce compared to the base alloy and La addition. Additives showed an increase of recalescence magnitude (TRAl-Si). Addition La and Ce increased the nucleation and growth temperature of Al-Cu phase. The microstructure analysis on the silicon morphology showed that 1 wt % La and 1 wt % Ce additions play refiner role in Al-Si-Cu near eutectic alloys. Findings are also confirmed by aspect ratio of eutectic silicon phase.

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47

Mihajlovic, Sanja, Silvia Lang, Marta V. Sut, Heimo Strohmaier, Christian J. Gruber, Günther Koraimann, Elena Cabezón, Gabriel Moncalián, Fernando de la Cruz, and Ellen L. Zechner. "Plasmid R1 Conjugative DNA Processing Is Regulated at the Coupling Protein Interface." Journal of Bacteriology 191, no. 22 (September 18, 2009): 6877–87. http://dx.doi.org/10.1128/jb.00918-09.

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ABSTRACT Selective substrate uptake controls initiation of macromolecular secretion by type IV secretion systems in gram-negative bacteria. Type IV coupling proteins (T4CPs) are essential, but the molecular mechanisms governing substrate entry to the translocation pathway remain obscure. We report a biochemical approach to reconstitute a regulatory interface between the plasmid R1 T4CP and the nucleoprotein relaxosome dedicated to the initiation stage of plasmid DNA processing and substrate presentation. The predicted cytosolic domain of T4CP TraD was purified in a predominantly monomeric form, and potential regulatory effects of this protein on catalytic activities exhibited by the relaxosome during transfer initiation were analyzed in vitro. TraDΔN130 stimulated the TraI DNA transesterase activity apparently via interactions on both the protein and the DNA levels. TraM, a protein interaction partner of TraD, also increased DNA transesterase activity in vitro. The mechanism may involve altered DNA conformation as TraM induced underwinding of oriT plasmid DNA in vivo (ΔLk = −4). Permanganate mapping of the positions of duplex melting due to relaxosome assembly with TraDΔN130 on supercoiled DNA in vitro confirmed localized unwinding at nic but ruled out formation of an open complex compatible with initiation of the TraI helicase activity. These data link relaxosome regulation to the T4CP and support the model that a committed step in the initiation of DNA export requires activation of TraI helicase loading or catalysis.

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Liu, L., H. Qi, J. Wang, and H. Lin. "PAPI, a novel TUDOR-domain protein, complexes with AGO3, ME31B and TRAL in the nuage to silence transposition." Development 138, no. 9 (March 29, 2011): 1863–73. http://dx.doi.org/10.1242/dev.059287.

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Audette, Gerald. "Structural, Functional and Dynamic Studies of F Plasmid T4SS Proteins." Acta Crystallographica Section A Foundations and Advances 70, a1 (August 5, 2014): C1285. http://dx.doi.org/10.1107/s2053273314087142.

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The transfer of genetic material within a bacterial population through the process of conjugation distributes novel genetic elements for survival in unique environments. Bacterial conjugation is important to public health as the spread antibiotic resistance genes among bacteria results in multi-drug resistance. Indeed, approximately 70% of bacteria that cause hospital-acquired infections are resistant to at least one antibiotic. Conjugative systems, such as the F plasmid of Escherichia coli, consist of proteins that share similarities to type IV secretion systems (T4SS). T4SS proteins of the F plasmid form a membrane spanning protein complex and surface exposed pilus. The periplasmic T4SS proteins TraF, TraW and TrbC play important roles during the F pilus assembly and DNA transfer. Functional analysis of a series of TraF mutants has shown that modification to TraF abolishes pilus synthesis and in turn F plasmid conjugation. In addition, dynamic analysis of TraF using time-resolved hydrogen-deuterium exchange mass spectrometry has revealed a well structured C-terminal thioredoxin-like domain and a more dynamic N-terminal domain that is predicted to interact with companion T4SS protein TraH. In addition, interaction analysis of the putative pore forming proteins TraW and TrbC indicate that the C-terminal domain of TrbC is not required for interaction with TraW, unlike previous models of F T4SS assembly. Rather the C-terminal domain of TraW preferentially interacts with the N-terminal domain of TrbC. These studies are providing a clearer picture of the structures and interactions that occur within the F T4SS assembly during the conjugative process.

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Su, Shengchang, Sharik R. Khan, and Stephen K. Farrand. "Induction and Loss of Ti Plasmid Conjugative Competence in Response to the Acyl-Homoserine Lactone Quorum-Sensing Signal." Journal of Bacteriology 190, no. 13 (January 18, 2008): 4398–407. http://dx.doi.org/10.1128/jb.01684-07.

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ABSTRACT Conjugative transfer of the Ti plasmids of Agrobacterium tumefaciens is controlled by a quorum-sensing system composed of TraR and its signal N-(3-oxo-octanoyl)-l-homoserine lactone. This system is, in turn, controlled by the conjugative opines produced by crown gall tumors induced on plants by the bacteria. Using nonpolar traI mutants, we examined the kinetics of induction of conjugative transfer in response to exogenous acyl-homoserine lactone. In the absence of the antiactivator TraM, onset of induction of transfer requires about 30 min, 15 to 20 min of which is needed for expression and construction of the conjugative apparatus. TraM delays the onset of conjugation by 30 min. While the rate of development of conjugative competence was not significantly affected by levels of TraR, maximum efficiencies of transfer were correlated with amounts of the activator in the donors. Donors harboring Ti plasmids lacking TraM were fully induced by the quormone at concentrations as low as 100 pM. TraM raised the concentration of signal required for maximum activity to 1 nM. Donors grown in batch culture retained conjugative competence following signal removal, even when in stationary phase. However, donors kept in balanced growth rapidly lost transfer ability following signal removal. Loss of transfer was mirrored by a decrease in levels of active TraR. Decreases in TraR activity and conjugative competence could be accounted for by dilution associated with cell division, suggesting that while induction of Ti plasmid conjugation is an active process, the cells lack a mechanism for disassembling the conjugative apparatus when signals become limiting.

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Journal articles on the topic 'Tral'

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