Characterization of the Tra2 Region of the IncHI1 Plasmid R27

Rooker, Michelle M.; Sherburne, Craig; Lawley, Trevor D.; Taylor, Diane E.

doi:10.1006/plas.1999.1396

Cited by 18 publications

(28 citation statements)

References 32 publications

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“…All ORFs are transcribed in the same direction, with the exception of ORF004 and TraO to TraZ, which appear to constitute an operon. Previous analysis indicated that the Tra2 region encodes mating-pair formation (Mpf) proteins that are related to Mpf proteins of IncF transfer systems, as determined by Basic Local Alignment Search Tool (BLAST) analysis (45). Prior to our mutational and functional analysis of the Tra2 region, we performed positionspecific iterated BLAST (PSI-BLAST) analysis, a more sensitive version of BLAST analysis, and protein alignments with the predicted transfer protein products of the Tra2 region and our findings are described quantitatively (Table 2).…”

Section: Resultsmentioning

confidence: 99%

“…Mutagenesis. Mutants with mutations of trhE, trhK, trhB, trhC, trhW, trhU, and trhN were created prior to this study using random mini-Tn10 mutagenesis, as described previously (45). Mini-Tn10 consists of a chloramphenicol resistance cassette flanked by Tn10 inverted repeats (23).…”

Section: Methodsmentioning

confidence: 99%

“…Seven mini-Tn10 transfer mutants have been identified within the Tra2 region (trhE, -K, -B, -C, -W, -U, and -N) (45). To identify a role in transfer for the remaining 17 Tra2 genes (the entire Tra2 excluding 4 partitioning genes), we systematically created gene disruptions of each of these ORFs in drR27, as previously described (26).…”

Section: Vol 185 2003 Conjugative Transfer Region 2 Of Inchi1 Plasmmentioning

confidence: 99%

“…drR27 is the prototypical IncHI1 plasmid that has an elevated transfer frequency compared to the wild type due to an insertion into htdA (48,52). In addition, we have performed a preliminary analysis of the Tra2 region (45). In this study, we performed a functional and mutational analysis of the Tra2 region and compared the essential transfer components of R27 to other transfer systems.…”

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confidence: 99%

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Functional and Mutational Analysis of Conjugative Transfer Region 2 (Tra2) from the IncHI1 Plasmid R27

Lawley¹,

Gilmour

Gunton

et al. 2003

J Bacteriol

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The transfer 2 region (Tra2) of the conjugative plasmid drR27 (derepressed R27) was analyzed by PSI-BLAST, insertional mutagenesis, genetic complementation, and an H-pilus assay. Tra2 contains 11 matingpair formation (Mpf) genes that are essential for conjugative transfer, 9 of which are essential for H-pilus production (trhA, -L, -E, -K, -B, -V, -C, -P, and -W). TrhK has similarity to secretin proteins, suggesting a mechanism by which DNA could traverse the outer membrane of donors. The remaining two Mpf genes, trhU and trhN, play an auxiliary role in H-pilus synthesis and are proposed to be involved in DNA transfer and mating-pair stabilization, respectively. Conjugative transfer abilities were restored for each mutant when complemented with the corresponding transfer gene. In addition to the essential Mpf genes, three genes, trhO, trhZ, and htdA, modulate R27 transfer frequency. Disruption of trhO and trhZ severely reduced the transfer frequencies of drR27, whereas disruption of htdA greatly increased the transfer frequency of wild-type R27 to drR27 levels. A comparison of the essential transfer genes encoded by the Tra2 and Tra1 (T.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Vol 185 2003 Conjugative Transfer Region 2 Of Inchi1 Plasmmentioning

confidence: 99%

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confidence: 99%

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Functional and Mutational Analysis of Conjugative Transfer Region 2 (Tra2) from the IncHI1 Plasmid R27

Lawley¹,

Gilmour

Gunton

et al. 2003

J Bacteriol

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“…The transfer genes of R27 are contained within two separate regions, designated Tra1 and Tra2. Since mutations in either transfer region abolish transfer, both transfer regions contribute to the R27 conjugative apparatus (28,30). An analysis of the Tra2 region has indicated that this region codes for mating pair formation and H-pilus synthesis phenotypes.…”

mentioning

confidence: 99%