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PMID: 7504666 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Temperature sensing in Yersinia pestis: translation of the LcrF activator protein is thermally regulated.

Journal of bacteriology ·Vol. 175 ·No. 24 ·1993-12-00 ·Pages 7901-9

Hoe NP, Goguen JD

Abstract

The lcrF gene of Yersinia pestis encodes a transcription activator responsible for inducing expression of several virulence-related proteins in response to temperature. The mechanism of this thermoregulation was investigated. An lcrF clone was found to produce much lower levels of LcrF protein at 26 than at 37 degrees C in Y. pestis, although it was transcribed at similar levels at both temperatures. High-level T7 polymerase-directed transcription of the lcrF gene in Escherichia coli also resulted in temperature-dependent production of the LcrF protein. Pulse-chase experiments showed that the LcrF protein was stable at 26 and 37 degrees C, suggesting that translation rate or message degradation is thermally controlled. The lcrF mRNA appears to be highly unstable and could not be reliably detected in Y. pestis. Insertion of the lcrF gene into plasmid pET4a, which produces high levels of plasmid-length RNA, aided detection of lcrF-specific message in E. coli. Comparison of the amount of LcrF protein produced per unit of message at 26 and 37 degrees C indicated that the efficiency of translation of lcrF message increased with temperature. mRNA secondary structure predictions suggest that the lcrF Shine-Dalgarno sequence is sequestered in a stem-loop. A model in which decreased stability of this stem-loop with increasing temperature leads to increased efficiency of translation initiation of lcrF message is presented.

Related Genes
lcrF
MeSH Terms
Bacterial Proteins/biosynthesis,isolation & purification Base Sequence DNA-Binding Proteins Gene Expression Regulation, Bacterial Genes, Bacterial Immunoblotting Molecular Sequence Data Nucleic Acid Conformation Protein Biosynthesis RNA, Bacterial/biosynthesis,chemistry,isolation & purification RNA, Messenger/biosynthesis,chemistry,isolation & purification Restriction Mapping Temperature Thermodynamics Trans-Activators Yersinia pestis/genetics,metabolism
Chemicals
Bacterial Proteins DNA-Binding Proteins RNA, Bacterial RNA, Messenger Trans-Activators lcrF protein, Yersinia pestis
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Hoe N P
Department of Molecular Genetics and Microbiology, University of Masschusetts Medical Center, Worcester 01655.
Goguen J D
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.175.24.7901-7909.1993
Published
1993-12-00
Pages
7901-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC206968
Subset
IM
Grants
NIAID NIH HHS · R01AI22176 · United States
External Links
PubMed source DOI: 10.1128/jb.175.24.7901-7909.1993 PMC Full Text View journal details
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