title: General secretion pathway (eps) genes required for toxin secretion and outer membrane biogenesis in Vibrio cholerae.
abstract: The general secretion pathway (GSP) of Vibrio cholerae is required for secretion of proteins including chitinase, enterotoxin, and protease through the outer membrane. In this study, we report the cloning and sequencing of a DNA fragment from V. cholerae, containing 12 open reading frames, epsC to -N, which are similar to GSP genes of Aeromonas, Erwinia, Klebsiella, Pseudomonas, and Xanthomonas spp. In addition to the two previously described genes, epsE and epsM (M. Sandkvist, V. Morales, and M. Bagdasarian, Gene 123: 81-86, 1993; L. J. Overbye, M. Sandkvist, and M. Bagdasarian, Gene 132:101-106, 1993), it is shown here that epsC, epsF, epsG, and epsL also encode proteins essential for GSP function. Mutations in the eps genes result in aberrant outer membrane protein profiles, which indicates that the GSP, or at least some of its components, is required not only for secretion of soluble proteins but also for proper outer membrane assembly. Several of the Eps proteins have been identified by use of the T7 polymerase-promoter system in Escherichia coli. One of them, a pilin-like protein, EpsG, was analyzed also in V. cholerae and found to migrate as two bands on polyacrylamide gels, suggesting that in this organism it might be processed or otherwise modified by a prepilin peptidase. We believe that TcpJ prepilin peptidase, which processes the subunit of the toxin-coregulated pilus, TcpA, is not involved in this event. This is supported by the observations that apparent processing of EpsG occurs in a tcpJ mutant of V. cholerae and that, when coexpressed in E. coli, TcpJ cannot process EpsG although the PilD peptidase from Neisseria gonorrhoeae can.
authors.affiliation: Oral Infection and Immunity Branch, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892-4350, USA.
authors.forename: M
authors.initials: M
authors.lastname: Sandkvist
authors.forename: L O
authors.initials: LO
authors.lastname: Michel
authors.forename: L P
authors.initials: LP
authors.lastname: Hough
authors.forename: V M
authors.initials: VM
authors.lastname: Morales
authors.forename: M
authors.initials: M
authors.lastname: Bagdasarian
authors.forename: M
authors.initials: M
authors.lastname: Koomey
authors.forename: V J
authors.initials: VJ
authors.lastname: DiRita
authors.forename: M
authors.initials: M
authors.lastname: Bagdasarian
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D036781:Fimbriae Proteins; D015964:Gene Expression Regulation, Bacterial; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D005810:Multigene Family; D016366:Open Reading Frames; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D013604:T-Phages / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism; D014734:Vibrio cholerae / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C079398:EpsE protein, Vibrio cholerae; C083759:EpsM protein, Vibrio cholerae; D008565:Membrane Proteins; D009097:Multienzyme Complexes; C085807:TcpA protein, Vibrio cholerae; D014157:Transcription Factors; C499324:toxR protein, Vibrio cholerae; C066719:toxR protein, bacteria; D036781:Fimbriae Proteins; D003851:Deoxyribonucleases; D010450:Endopeptidases; C092637:Tcpj protein, Vibrio cholerae