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PMID: 1624422 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Temperature sensing in Yersinia pestis: regulation of yopE transcription by lcrF.

Journal of bacteriology ·Vol. 174 ·No. 13 ·1992-07-00 ·Pages 4275-86

Hoe NP, Minion FC, Goguen JD

Abstract

In Escherichia coli, a yopE::lacZ fusion was found to be regulated by temperature in the presence of the cloned BamHI G fragment of Yersinia pestis plasmid pCD1, which contains the lcrF locus. Increasing the copy number of lcrF relative to that of the yopE reporter had a negligible effect on the induction ratio (26 versus 37 degrees C) but caused large reductions in the absolute levels of yopE transcription. We localized the lcrF gene by monitoring the induction phenotype of BamHI G deletion derivatives. Sequencing revealed an open reading frame capable of encoding a protein of 30.8 kDa. A protein product of this size was detected in a T7 expression system, and LcrF-dependent yopE-specific DNA binding activity was observed. As expected, LcrF exhibited 98% homology to VirF of Yersinia enterocolitica and significant homology to the carboxy termini of other members of the AraC family of transcriptional regulatory proteins. These proteins could be divided into two classes according to function: those regulating operons involved in catabolism of carbon and energy sources and those involved in regulating virulence genes. lcrF::lacZ transcriptional fusions were constructed and analyzed in Y. pestis and E. coli. The activity of the fusions was not affected by the native pCD1 virulence plasmid, an intact lcrF gene, or temperature. Thus, induction of lcrF transcription is not essential for temperature-dependent activation of yopE transcription. A portion of LcrF was found associated with the membrane fraction in E. coli; however, pulse-chase experiments indicated that this result is an artifact of fractionation.

Related Genes
lcrF yopE
MeSH Terms
Amino Acid Sequence Bacteria/genetics Bacterial Outer Membrane Proteins/genetics,metabolism Bacterial Proteins/genetics,metabolism Chromosome Deletion Cloning, Molecular/methods DNA, Bacterial/genetics,isolation & purification DNA-Binding Proteins Escherichia coli/genetics,physiology Gene Expression Regulation, Bacterial Genes, Bacterial Genotype Leucine Zippers/genetics Molecular Sequence Data Molecular Weight Open Reading Frames Plasmids Recombinant Fusion Proteins/metabolism Restriction Mapping Sequence Homology, Nucleic Acid Temperature Trans-Activators Transcription, Genetic Yersinia pestis/genetics,physiology beta-Galactosidase/genetics,metabolism
Chemicals
Bacterial Outer Membrane Proteins Bacterial Proteins DNA, Bacterial DNA-Binding Proteins Recombinant Fusion Proteins Trans-Activators lcrF protein, Yersinia pestis yopE protein, Yersinia beta-Galactosidase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hoe N P
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical Center, Worcester 01655.
Minion F C
Goguen J D
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.174.13.4275-4286.1992
Published
1992-07-00
Pages
4275-86
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC206210
Subset
IM
Grants
NIAID NIH HHS · AI22176 · United States
Databases
GENBANK
M79367, M79368, M79369, M79370, M79371, M79372, M84980, M8669, M86690, M91199, Z11493
External Links
PubMed source DOI: 10.1128/jb.174.13.4275-4286.1992 PMC Full Text View journal details
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