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Molecular analysis of lcrGVH, the V antigen operon of Yersinia pestis.

ABSTRACT: The lcrGVH operon of plasmid pCD1 in Yersinia pestis KIM encodes the virulence-associated V antigen, the regulatory protein LcrH, and LcrG, a protein of undefined function. In this study we sequenced lcrGVH and analyzed it for transcription initiation sites. There were three open reading frames within the sequence, 288, 981, and 507 bases in length, which could encode proteins with molecular weights and isoelectric points corresponding to those of LcrG, LcrV (V antigen), and LcrH, respectively. The predicted LcrV protein lacked an N-terminal signal sequence; however, an internal signallike sequence was present. An Escherichia coli-like promoter consensus sequence was detected upstream from lcrG. Primer extension analysis showed that (i) the transcriptional start site for lcrGVH was spaced only three bases upstream from the nearest ATG potential start site, raising the possibility that Y. pestis may use an alternate initiation codon for the V operon; (ii) there was much more primer-extended product in yersiniae grown in the absence of Ca2+ than in its presence, showing for the first time that lcrGVH is regulated at the transcriptional level by Ca2+; (iii) no separate lcrV initiation was detected, indicating that the V antigen is expressed from messages initiating at lcrG; and (iv) a non-Ca2+-regulated transcriptional start site was found upstream from lcrH, suggesting that the LcrH protein is expressed constitutively. However, two-dimensional gel analysis showed that net LcrH expression was regulated by Ca2+. We propose that lcrH lies within two differentially regulated operons, its own and lcrGVH.

SUBMITTER: Price SB

PROVIDER: S-EPMC210409 | biostudies-other | 1989 Oct

REPOSITORIES: biostudies-other

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Molecular analysis of lcrGVH, the V antigen operon of Yersinia pestis.

Price S B SB Leung K Y KY Barve S S SS Straley S C SC

Journal of bacteriology 19891001 10

The lcrGVH operon of plasmid pCD1 in Yersinia pestis KIM encodes the virulence-associated V antigen, the regulatory protein LcrH, and LcrG, a protein of undefined function. In this study we sequenced lcrGVH and analyzed it for transcription initiation sites. There were three open reading frames within the sequence, 288, 981, and 507 bases in length, which could encode proteins with molecular weights and isoelectric points corresponding to those of LcrG, LcrV (V antigen), and LcrH, respectively. ...[more]

PMID: 2477361

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Project description:BACKGROUND:Yersinia pestis initiates infection by parasitism of host macrophages. In response to macrophage infections, intracellular Y. pestis can assume a filamentous cellular morphology which may mediate resistance to host cell innate immune responses. We previously observed the expression of Y. pestis tellurite resistance proteins TerD and TerE from the terZABCDE operon during macrophage infections. Others have observed a filamentous response associated with expression of tellurite resistance operon in Escherichia coli exposed to tellurite. Therefore, in this study we examine the potential role of Y. pestis tellurite resistance operon in filamentous cellular morphology during macrophage infections. PRINCIPAL FINDINGS:In vitro treatment of Y. pestis culture with sodium tellurite (Na2TeO3) caused the bacterial cells to assume a filamentous phenotype similar to the filamentous phenotype observed during macrophage infections. A deletion mutant for genes terZAB abolished the filamentous morphologic response to tellurite exposure or intracellular parasitism, but without affecting tellurite resistance. However, a terZABCDE deletion mutant abolished both filamentous morphologic response and tellurite resistance. Complementation of the terZABCDE deletion mutant with terCDE, but not terZAB, partially restored tellurite resistance. When the terZABCDE deletion mutant was complemented with terZAB or terCDE, Y. pestis exhibited filamentous morphology during macrophage infections as well as while these complemented genes were being expressed under an in vitro condition. Further in E. coli, expression of Y. pestis terZAB, but not terCDE, conferred a filamentous phenotype. CONCLUSIONS:These findings support the role of Y. pestis terZAB mediation of the filamentous response phenotype; whereas, terCDE confers tellurite resistance. Although the beneficial role of filamentous morphological responses by Y. pestis during macrophage infections is yet to be fully defined, it may be a bacterial adaptive strategy to macrophage associated stresses.

Dataset Information

Molecular analysis of lcrGVH, the V antigen operon of Yersinia pestis.

Publications

Molecular analysis of lcrGVH, the V antigen operon of Yersinia pestis.

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