Transcript analysis of the plasmid R100 traJ and finP genes

Dempsey, Walter B.

doi:10.1007/bf00331160

Cited by 37 publications

(41 citation statements)

References 40 publications

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“…Analysis of traJ transcripts by Northern (RNA) blots showed similar amounts but different sizes of R100 traJ transcripts in repressed (RlOOfinP+ finO+) and derepressed (R100-lfinP+ finO) strains (2). In these analyses, R100 traJ RNA was seen as a collection of aborted transcripts much like the lac mRNA seen by Stanssens et al (13) when lac translation was blocked.…”

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confidence: 62%

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Derepression of conjugal transfer of the antibiotic resistance plasmid R100 by antisense RNA

Dempsey¹

1989

J Bacteriol

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confidence: 62%

“…traJ protein is required for transcription of the 30-kilobase-long transfer operon (15), finP has been proposed to be an antisense RNA that blocks ribosome binding to the traJ transcript (4,6,11), and finO has been proposed to be a small RNA that maximizes correct folding of the traJ transcript during its reaction with finP (2).…”

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confidence: 99%

Derepression of conjugal transfer of the antibiotic resistance plasmid R100 by antisense RNA

Dempsey¹

1989

J Bacteriol

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“…In the finO mutant R100-1, transcription across the traJ leader is much greater than it is in R100, and full-length transcripts can be detected (Dempsey, 1987). In R100, the finP gene is much more heavily transcribed than it is in R100-1 (Dempsey, 1987). Northern blots show many more finP transcripts than traJ leader transcripts in R100-1 (Dempsey, 1994a), and the difference is more pronounced in R100 (W. B. Dempsey, unpublished observation).…”

Section: R100 (Nr1 R222mentioning

confidence: 99%

“…finP and the untranslated leader region of traJ are transcribed continuously in both R100 and R100-1 (Dempsey, 1987). In R100, small amounts of traJ leader transcript are detected on Northern blots, but full-length traJ transcripts are not (Dempsey, 1987).…”

Section: R100 (Nr1 R222mentioning

confidence: 99%

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The finM promoter and the traM promoter are the principal promoters of the traM gene of the antibiotic resistance plasmid R100

Stockwell

Dempsey

1997

Molecular Microbiology

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SummaryfinP multicopy repression and traJ multicopy derepression indicate that the ratio of sense to antisense transcripts is important in the regulation of R100 conjugation. The extension of R100 traM transcripts into traJ shows that promoters in traM can affect this ratio, making the regulation of traM transcription important in the regulation of R100 conjugation. Since R100 traM, traY and traI proteins bind to the traM promoter region, we examined traM transcription in R100-1 traM, traY and traI mutants and compared it with traM transcription in both R100-1 and R100. We verified that the traM and finM promoters provide virtually all the transcripts originating in the R100-1 traM gene. When either is deleted, as in VAR22 or VAR30, the remaining promoter is highly active. We show here that traY positively regulates R100-1 traM transcription, as has been found for F. We found that traI did not regulate R100-1 traM transcription. The measured activity of the native R100 traM promoter was 12% of that in R100-1, whereas the native R100 finM promoter was 45% of that in R100-1. These data and data from the R100-1 traY and traI mutants show that the activities of the two promoters varied independently.

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Control of replication of plasmid R1: kinetics of in vitro interaction between the antisense RNA, CopA, and its target, CopT.

1988

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The frequency of replication of IncFII plasmids is regulated by the availability of a rate‐limiting protein, RepA. This protein acts to promote initiation of replication and its synthesis is negatively controlled both at the transcriptional and translational level. The translational control is exerted by the binding of a small antisense RNA, CopA RNA, to its target, CopT, which is located in the leader region of the RepA mRNA. As a consequence, formation of RepA is inhibited. Here we demonstrate the binding of CopA RNA to CopT RNA in vitro; the rate constant of binding was determined to be approximately 1 X 10(6) M‐1 s‐1 at 37 degrees C. We have also shown that in vitro synthesized RepA mRNA molecules differing in length, but which contain the whole CopT region, are able to bind CopA RNA with similar rates. Analysis of the binding of CopA/CopT molecules derived from a copy‐number mutant plasmid showed that the effect of the mutation on the rate of in vitro binding correlates well with its phenotypic effect in vivo, i.e. the binding rate constant is lowered in proportion to the increase in copy number. Likewise, the result of an in vitro incompatibility test is in agreement with in vivo data.

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Transcript analysis of the plasmid R100 traJ and finP genes

Cited by 37 publications

References 40 publications

Derepression of conjugal transfer of the antibiotic resistance plasmid R100 by antisense RNA

Derepression of conjugal transfer of the antibiotic resistance plasmid R100 by antisense RNA

The finM promoter and the traM promoter are the principal promoters of the traM gene of the antibiotic resistance plasmid R100

Control of replication of plasmid R1: kinetics of in vitro interaction between the antisense RNA, CopA, and its target, CopT.

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