Functional and Mutational Analysis of Conjugative Transfer Region 1 (Tra1) from the IncHI1 Plasmid R27

Lawley, Trevor D.; Gilmour, Matthew W.; Gunton, James E.; Standeven, Leah J.; Taylor, Diane E.

doi:10.1128/jb.184.8.2173-2180.2002

Cited by 59 publications

(79 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although we know of no molecular reason that can explain this difference, we speculate that large plasmids might need additional regulation. Accordingly, MOB F and MOB H relaxosomes are structurally more complex and contain comparably large oriTs (Ͼ300 bp) (2,51,81,92,118). On the other hand, MOB Q1 and certain MOB V relaxosomes are simple, contain short oriTs (Ͻ100 bp), and are promiscuous in their use of different helper T4SSs as mating channels (49,52,100).…”

Section: Mobility Of Genbank Plasmidsmentioning

confidence: 99%

Mobility of Plasmids

Smillie

Garcillán-Barcia

Francia

et al. 2010

Microbiol Mol Biol Rev

981

1,054

View full text Add to dashboard Cite

International audiencePlasmids are key vectors of horizontal gene transfer and essential genetic engineering tools. They code for genes involved in many aspects of microbial biology, including detoxication, virulence, ecological interactions, and antibiotic resistance. While many studies have decorticated the mechanisms of mobility in model plasmids, the identification and characterization of plasmid mobility from genome data are unexplored. By reviewing the available data and literature, we established a computational protocol to identify and classify conjugation and mobilization genetic modules in 1,730 plasmids. This allowed the accurate classification of proteobacterial conjugative or mobilizable systems in a combination of four mating pair formation and six relaxase families. The available evidence suggests that half of the plasmids are nonmobilizable and that half of the remaining plasmids are conjugative. Some conjugative systems are much more abundant than others and preferably associated with some clades or plasmid sizes. Most very large plasmids are nonmobilizable, with evidence of ongoing domestication into secondary chromosomes. The evolution of conjugation elements shows ancient divergence between mobility systems, with relaxases and type IV coupling proteins (T4CPs) often following separate paths from type IV secretion systems. Phylogenetic patterns of mobility proteins are consistent with the phylogeny of the host prokaryotes, suggesting that plasmid mobility is in general circumscribed within large clades. Our survey suggests the existence of unsuspected new relaxases in archaea and new conjugation systems in cyanobacteria and actinobacteria. Few genes, e.g., T4CPs, relaxases, and VirB4, are at the core of plasmid conjugation, and together with accessory genes, they have evolved into specific systems adapted to specific physiological and ecological contexts

show abstract

Section: Mobility Of Genbank Plasmidsmentioning

confidence: 99%

Mobility of Plasmids

Smillie

Garcillán-Barcia

Francia

et al. 2010

Microbiol Mol Biol Rev

981

1,054

View full text Add to dashboard Cite

show abstract

“…Conjugal transfer of drR27 was performed as previously described (26). For complementation experiments, E. coli donors (DY330R) which contained the R27 Tra2 mutant and an expression vector encoding the appropriate transfer protein were mated with recipients (DY330N).…”

Section: Methodsmentioning

confidence: 99%

“…NC_002305). All remaining Tra2 genes (trhA, trhL, orf030, orf028, orf027, trhV, trhZ, orf017, orf016, trhO, htdA, htdF, htdK, orf009, trhP, orf004, and trhI) were mutated by gene disruption using the E. coli recombination system as described previously (26,53). Gene disruptions were created by insertion of a chloramphenicol resistance cassette (cat from mini-Tn10) into each of the above-mentioned ORFs in a sequence-specific fashion.…”

Section: Methodsmentioning

confidence: 99%

“…To determine the Hgal infectivity of Tra2 mutants, phage spot tests were performed as previously described (26,31).…”

Section: Methodsmentioning

confidence: 99%

“…The continued prevalence of IncHI1 plasmids can be partially attributed to the conjugative transfer functions of these plasmids, since horizontal transfer maintains a plasmid within a bacterial population. We have previously characterized the Tra1 region of derepressed R27 (drR27) and found that it contains the origin of transfer and genes encoding the relaxosome, coupling protein, and three Mpf proteins (26). 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).…”

mentioning

confidence: 99%

See 2 more Smart Citations

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

Lawley¹,

Gilmour

Gunton

et al. 2003

J Bacteriol

Self Cite

View full text Add to dashboard Cite

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.

show abstract

HDX‐MS and deletion analysis of the type 4 secretion system protein TraF from the Escherichia coli F plasmid

et al. 2016

View full text Add to dashboard Cite

Conjugative DNA transfer by the F‐plasmid is achieved through a type IV secretion system (T4SS) encoded within the plasmid's transfer region; TraF is one of several F‐T4SS proteins essential for F‐pilus assembly. In order to identify regions of the protein important for TraF function, a series of deletion mutants were assessed for their ability to recover conjugative transfer in a traF knockout. Interestingly, modification of any region of TraF abolishes pilus synthesis, resulting in a loss of rescue of conjugative function. Dynamic analysis of TraF by time‐resolved hydrogen–deuterium exchange revealed that the C‐terminal region containing the predicted thioredoxin‐like domain is quite structured, while the N‐terminal region, predicted to interact with TraH in the intact F‐T4SS, was more dynamic.

show abstract

Functional and Mutational Analysis of Conjugative Transfer Region 1 (Tra1) from the IncHI1 Plasmid R27

Cited by 59 publications

References 35 publications

Mobility of Plasmids

Mobility of Plasmids

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

HDX‐MS and deletion analysis of the type 4 secretion system protein TraF from the Escherichia coli F plasmid

Contact Info

Product

Resources

About