Molecular and functional characterization of the Salmonella typhimurium invasion genes invB and invC: homology of InvC to the F0F1 ATPase family of proteins.
Entry into intestinal epithelial cells is an essential step in the pathogenesis of Salmonella infections. Our laboratory has previously identified a genetic locus, inv, that is necessary for efficient entry of Salmonella typhimurium into cultured epithelial cells. We have carried out a molecular and functional analysis of invB and invC, two members of this locus. The nucleotide sequence of these genes indicated that invB and invC encode polypeptides with molecular masses of 15 and 47 kDa, respectively. Polypeptides with the predicted sizes were observed when these genes were expressed under the control of a T7 promoter. Strains carrying nonpolar mutations in these genes were constructed, and their phenotypes were examined in a variety of assays. A mutation in invC rendered S. typhimurium defective in their ability to enter cultured epithelial cells, while mutations in invB did not. Comparison of the predicted sequences of InvB and InvC with translated sequences in GenBank revealed that these polypeptides are similar to the Shigella spp. proteins Spa15 and Spa47, which are involved in the surface presentation of the invasion protein antigens (Ipa) of these organisms. In addition, InvC showed significant similarity to a protein family which shares sequence homology with the catalytic beta subunit of the F0F1 ATPase from a number of microorganisms. Consistent with this finding, purified preparations of InvC showed significant ATPase activity. Site-directed mutagenesis of a residue essential for the catalytical function of this family of proteins resulted in a protein devoid of ATPase activity and unable to complement an invC mutant of S. typhimurium. These results suggest that InvC may energize the protein export apparatus encoded in the inv locus which is required for the surface presentation of determinants needed for the entry of Salmonella species into mammalian cells. The role of InvB in this process remains uncertain.
Eichelberg
K
K
Department of Microbiology, School of Medicine, SUNY Stony Brook 11794-5222.
Ginocchio
C C
CC
Galán
J E
JE
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D015870:Gene Expression; D005798:Genes, Bacterial / Q000235:genetics*; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D011485:Protein Binding; D006180:Proton-Translocating ATPases / Q000235:genetics*; D012486:Salmonella typhimurium / Q000235:genetics* / Q000472:pathogenicity; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014774:Virulence / Q000235:genetics
D001426:Bacterial Proteins; C088597:invB protein, Salmonella; C088598:invC protein, Salmonella typhimurium; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases