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PMID: 9696755 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Both the fipA gene of pKM101 and the pifC gene of F inhibit conjugal transfer of RP1 by an effect on traG.

Journal of bacteriology ·Vol. 180 ·No. 16 ·1998-08-00 ·Pages 4093-101

Santini JM, Stanisich VA

Abstract

The mechanisms by which gene products inhibit the conjugal transfer of IncP plasmids (e.g., RP1) have been little studied. We have isolated and characterized one such gene, fipA (624 nucleotides), from the SmaI (14.8 kb)-AatII (15.6 kb) region of pKM101(IncN). This gene, which is also conserved in other IncN plasmids, is transcribed in an anticlockwise direction, probably as part of a transfer operon that includes traHI. The FipA protein (24 kDa) appears to be cytoplasmic and, when expressed from a multicopy plasmid, retards the growth of Escherichia coli WP2. The mode of action of fipA was compared with that of the apparently unrelated pifC gene from F(IncFI). Both genes inhibit the transfer of IncPalpha and IncPbeta plasmids but to different degrees. They also inhibit the mobilization of RSF1010 (which requires the RP1 pilus genes and traG) but not of CloDF13 (which encodes a traG homolog). Evidence that traG was the specific target of inhibition was obtained in an artificial system in which cloned traG was used to enhance RSF1010 mobilization via the N pilus system. Such enhancement did not occur in the presence of fipA or pifC. The availability of an in vivo assay of PifC enabled us to show that F pif operon expression increased in cells carrying F'lac and traG, but only if the traG coding sequence was intact. This finding suggested that conjugal inhibition of RP1 was most likely due to a PifC-TraG protein interaction. On phenotypic grounds inhibition of traG by fipA is also likely to occur posttranscriptionally. Whether or not the selection of traG as the inhibition target is an evolutionary tactic to limit the spread of P plasmids, we anticipate that fipA and pifC will prove useful in further investigation of the conjugal roles of traG and its homologs.

MeSH Terms
Bacterial Proteins/genetics Cloning, Molecular Conjugation, Genetic Escherichia coli/genetics,metabolism Escherichia coli Proteins Eukaryotic Initiation Factor-3 F Factor/genetics Genes, Bacterial Membrane Proteins Peptide Initiation Factors/genetics,metabolism R Factors/genetics
Chemicals
Bacterial Proteins Escherichia coli Proteins Eukaryotic Initiation Factor-3 Membrane Proteins Peptide Initiation Factors TraG protein, E coli
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Santini J M
Department of Microbiology, La Trobe University, Bundoora 3083, Australia.
Stanisich V A
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.180.16.4093-4101.1998
Published
1998-08-00
Pages
4093-101
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC107403
Subset
IM
Databases
GENBANK
U42978
External Links
PubMed source DOI: 10.1128/JB.180.16.4093-4101.1998 PMC Full Text View journal details
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