Molecular characterization of a trans-acting, positive effector (ipaR) of invasion plasmid antigen synthesis in Shigella flexneri serotype 5.
A trans-acting, positive effector of invasion plasmid antigen (Ipa) synthesis has been identified and mapped on the pWR100 invasion plasmid of Shigella flexneri serotype 5 (strain M90T-W). Recombinant plasmids carrying this regulatory gene, designated ipaR, were found to restore full virulence to a non-invasive ipaR::Tn5 insertion mutant [M90T-W(pHS1042)] that had lost the ability to synthesize four Ipa antigens (IpaA, 70 kDa; IpaB, 62 kDa; IpaC, 42 kDa; and IpaD, 37 kDa). Genetic mapping of the ipaR gene positioned the locus on a 2.6 kb PstI-AccI fragment contained within a larger 8.0 kb EcoRI molecule that also encoded IpaD, IpaA, and two small proteins (27 kDa and 28 kDa). The trans regulatory effect of the ipaR product on ipaB, ipaC, ipaD, and ipaA expression was demonstrated by transforming compatible ipaBC, ipaDA, ipaR and ipaDAR plasmid recombinants, in various combinations, into M90T-A3, an isogenic invasion plasmid mutant of M90T-W that contained a deletion of the pWR100 ipaBCDA and ipaR loci; such transformants produced wild type levels of the IpaB, IpaC, IpaD and IpaA antigens only in the presence of IpaR+ plasmids. DNA sequence analysis of the ipaR region established that the intiation codon for ipaR is 459 bp from the 3'-end of the ipaA gene and that ipaR encodes a 309 amino acid residue protein. An interesting feature of the IpaR polypeptide was its strong sequence homology with the bacteriophage P1 partition protein ParB, consisting of a 42.8% amino acid identity over a 278 residue section of the aligned proteins.
Buysse
J M
JM
Department of Bacterial Immunology, Walter Reed Army Institute of Research, Walter Reed Army Medical Center, Washington, DC 20307-5100.
Venkatesan
M
M
Mills
J A
JA
Oaks
E V
EV
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology; D001426:Bacterial Proteins; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D005809:Genes, Regulator; D005816:Genetic Complementation Test; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D012763:Shigella flexneri / Q000235:genetics / Q000276:immunology*; D015534:Trans-Activators / Q000235:genetics*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; C063664:IpaA protein, Shigella flexneri; D015534:Trans-Activators