Abstract |
A random library of Escherichia coli MG1655 genomic fragments fused to a promoterless green fluorescent protein (GFP) gene was constructed and screened by differential fluorescence induction for promoters that are induced after exposure to a sublethal high hydrostatic pressure stress. This screening yielded three promoters of genes belonging to the heat shock regulon (dnaK, lon, clpPX), suggesting a role for heat shock proteins in protection against, and/or repair of, damage caused by high pressure. Several further observations provide additional support for this hypothesis: (i). the expression of rpoH, encoding the heat shock-specific sigma factor sigma(32), was also induced by high pressure; (ii). heat shock rendered E. coli significantly more resistant to subsequent high-pressure inactivation, and this heat shock-induced pressure resistance followed the same time course as the induction of heat shock genes; (iii). basal expression levels of GFP from heat shock promoters, and expression of several heat shock proteins as determined by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins extracted from pulse-labeled cells, was increased in three previously isolated pressure-resistant mutants of E. coli compared to wild-type levels.
|
Authors | Abram Aertsen, Kristof Vanoirbeek, Philipp De Spiegeleer, Jan Sermon, Kristel Hauben, Anne Farewell, Thomas Nyström, Chris W Michiels |
Journal | Applied and environmental microbiology
(Appl Environ Microbiol)
Vol. 70
Issue 5
Pg. 2660-6
(May 2004)
ISSN: 0099-2240 [Print] United States |
PMID | 15128516
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
|
Chemical References |
- Culture Media
- Escherichia coli Proteins
- Heat-Shock Proteins
- Luminescent Proteins
- Green Fluorescent Proteins
|
Topics |
- Culture Media
- Escherichia coli
(genetics, growth & development)
- Escherichia coli Proteins
(genetics, metabolism)
- Gene Expression Regulation, Bacterial
- Green Fluorescent Proteins
- Heat-Shock Proteins
(genetics, metabolism)
- Heat-Shock Response
- Hydrostatic Pressure
- Luminescent Proteins
(genetics, metabolism)
- Promoter Regions, Genetic
|