Identification of PhoP-PhoQ activated genes within a duplicated region of the Salmonella typhimurium chromosome.
Salmonellae virulence requires the PhoP-PhoQ two-component regulatory system. PhoP-PhoQ activate the transcription of genes following phagocytosis by macrophages which are necessary for survival within the phagosome environment. Thirteen previously undefined PhoP-activated gene fusions generated by MudJ and TnphoA (pag A, and E-P, respectively) were cloned and sequenced. Most pag products show no similarity to proteins in the database, while others are predicted to encode: a UDP-glucose dehydrogenase (pagA); a protein with similarity to the product of an E. coli aluminium-induced gene (pagH); a protein encoded within a Salmonella-unique region adjacent to the sinR gene (pagN); a protein similar to a product of the Yersinia virulence plasmid (pagO); and a protein with similarity to CrcA which is necessary for resistance of E. coli to camphor (pagP). Of the pag characterized, only pagK, M and O were closely linked. pagJ and pagK were shown to be unlinked but nearly identical in DNA sequence, as each was located within a 1.6 kb DNA duplication. The translations of sequences surrounding pagJ and pagK show similarity to proteins from extrachromosomal elements as well as those involved in DNA transposition and rearrangement, suggesting that this region may have been or is a mobile element. The transcriptional start sites of pagK, M, and J were determined; however, comparison to other known pag gene promoters failed to reveal a consensus sequence for PhoP-regulated activation. DNA sequences hybridizing to a Salmonella typhimurium pagK specific probe were found in S. enteritidis but absent in other Salmonella serotypes and Enterobacteriaceae tested, suggesting that these genes are specific for broad host range Salmonellae that cause diarrhoea in humans. Cumulatively, these data further demonstrate: (1) that PhoP-PhoQ is a global regulator of the production of diverse envelope or secreted proteins; (2) that PhoP-PhoQ regulate the production of proteins of redundant function; and (3) that pag are often located in regions of horizontally acquired DNA that are absent in other Enterobacteriaceae.
Department of Medicine, University of Washington, HSB K-140, Box 357710, Seattle, WA 98195, USA.
J S
JS
Gunn
W J
WJ
Belden
S I
SI
Miller
D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000378:metabolism*; D002872:Chromosome Deletion; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D005819:Genetic Markers; D015698:Genomic Library; D008969:Molecular Sequence Data; D005810:Multigene Family; D016296:Mutagenesis; D012486:Salmonella typhimurium / Q000235:genetics* / Q000472:pathogenicity*; D014157:Transcription Factors / Q000378:metabolism; D014774:Virulence / Q000235:genetics
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D005819:Genetic Markers; C079792:PhoQ protein, Bacteria; D014157:Transcription Factors; C061064:PhoP protein, Bacteria