Analysis of the traLEKBP sequence and the TraP protein from three F-like plasmids: F, R100-1 and ColB2

Anthony, Karen G.; Kathir, Pushpa; Moore, Deanna; Ippen‐Ihler, Karin; Frost, Laura S.

doi:10.1128/jb.178.11.3194-3200.1996

Cited by 18 publications

(11 citation statements)

References 26 publications

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“…e The data on traP is taken from Anthony et al (4). traP mutants have fragile pili that appear to fall off the cell easily during preparation for electron microscopy (unpublished observation).…”

Section: Discussionmentioning

confidence: 99%

Comparison of Proteins Involved in Pilus Synthesis and Mating Pair Stabilization from the Related Plasmids F and R100-1: Insights into the Mechanism of Conjugation

et al. 1999

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F and R100-1 are closely related, derepressed, conjugative plasmids from the IncFI and IncFII incompatibility groups, respectively. Heteroduplex mapping and genetic analyses have revealed that the transfer regions are extremely similar between the two plasmids. Plasmid specificity can occur at the level of relaxosome formation, regulation, and surface exclusion between the two transfer systems. There are also differences in pilus serology, pilus-specific phage sensitivity, and requirements for OmpA and lipopolysaccharide components in the recipient cell. These phenotypic differences were exploited in this study to yield new information about the mechanism of pilus synthesis, mating pair stabilization, and surface and/or entry exclusion, which are collectively involved in mating pair formation (Mpf). The sequence of the remainder of the transfer region of R100-1 (trbA to traS) has been completed, and the complete sequence is compared to that of F. The differences between the two transfer regions include insertions and deletions, gene duplications, and mosaicism within genes, although the genes essential for Mpf are conserved in both plasmids. F+ cells carrying defined mutations in each of the Mpf genes were complemented with the homologous genes from R100-1. Our results indicate that the specificity in recipient cell recognition and entry exclusion are mediated by TraN and TraG, respectively, and not by the pilus.

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Section: Discussionmentioning

confidence: 99%

Comparison of Proteins Involved in Pilus Synthesis and Mating Pair Stabilization from the Related Plasmids F and R100-1: Insights into the Mechanism of Conjugation

et al. 1999

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“…Plasmids pGAP60 (Pogliano et al ., 2001), pGAP40 (Brendler et al ., 2000) and pOX38kan (Anthony et al ., 1996) were described previously. Plasmid pGAP500 was constructed by cloning the 10 kb lacO array from plasmid pAFS52 (Straight et al ., 1996) into pRR10 (Roberts et al ., 1990).…”

Section: Methodsmentioning

confidence: 99%

Intracellular mobility of plasmid DNA is limited by the ParA family of partitioning systems

Derman

Lim-Fong

Pogliano³

2008

Molecular Microbiology

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SummaryThe highly conserved ParA family of partitioning systems is responsible for positioning DNA and protein complexes in bacteria. In Escherichia coli, plasmids that rely upon these systems are positioned at mid-cell and are repositioned at the quarter-cell positions after replication. How they remain fixed at these positions throughout the cell cycle is unknown. We use fluorescence recovery after photobleaching and time-lapse microscopy to measure plasmid mobility in living E. coli cells. We find that a minimalized version of plasmid RK2 that lacks its Par system is highly mobile, that the intact RK2 plasmid is relatively immobile, and that the addition of a Par system to the minimalized RK2 plasmid limits its mobility to that of the intact RK2. Mobility is thus the default state, and Par systems are required not only to position plasmids, but also to hold them at these positions. The intervention of Par systems is required continuously throughout the cell cycle to restrict plasmid movement that would, if unrestricted, subvert the segregation process. Our results reveal an important function for Par systems in plasmid DNA segregation that is likely to be conserved in bacteria.

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“…Samples (3 ml) were removed for both RNA and DNA isolation. The presence of pOX38-Km in the various strains was occasionally confirmed by checking for phage sensitivity using a spot test (Anthony et al, 1996).…”

Section: Methodsmentioning

confidence: 99%

F- phenocopies: characterization of expression of the F transfer region in stationary phase

Frost

Manchak

1998

Microbiology

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The phenomenon of 'F' phenocopies' in which F+ cells become transferdeficient in stationary phase seems contradictory to the proposed role for F transfer in adaptive mutation during stationary phase induced by nutrient limitation. The expression of a range of transfer genes a t the transcriptional and translational level in stationary phase has been characterized as well as the degree of nicking at the origin of transfer, oriT. Transfer efficiency rapidly decreased in mid-exponential phase, coincident with a decrease in traM transcripts. Approximately 2 h later, the transcript for traA, encoding F-pilin, also decreased t o undetectable levels. The levels of TraA (pilin), TraD, Tral and TraT remained fairly constant well into stationary phase while the levels of TraM and Tral decreased t o undetectable levels in early stationary phase. A null mutation in the gene for the alternative G factor, rpoS, did not affect mating efficiency or transcript levels but did increase the stability of TraM and Tral in stationary phase. Nicking at oriT was detected a t maximal levels in early stationary phase and at low levels in late stationary phase. The results suggest that the F-pilus transfer apparatus is maintained in the cell envelope after transcription of the transfer region from the main promoter, Py, has ceased with down-regulation of traM transcription being the first step detected in this process. The presence of a low level of nicking a t oriT in stationary phase is consistent with a role for F in promoting adaptive mutation.

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Analysis of the traLEKBP sequence and the TraP protein from three F-like plasmids: F, R100-1 and ColB2

Cited by 18 publications

References 26 publications

Comparison of Proteins Involved in Pilus Synthesis and Mating Pair Stabilization from the Related Plasmids F and R100-1: Insights into the Mechanism of Conjugation

Comparison of Proteins Involved in Pilus Synthesis and Mating Pair Stabilization from the Related Plasmids F and R100-1: Insights into the Mechanism of Conjugation

Intracellular mobility of plasmid DNA is limited by the ParA family of partitioning systems

F- phenocopies: characterization of expression of the F transfer region in stationary phase

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