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Applied and Environmental Microbiology, May 2004, p. 2660-2666, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.2660-2666.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Heat Shock Protein-Mediated Resistance to High Hydrostatic Pressure in Escherichia coli

Abram Aertsen,1 Kristof Vanoirbeek,1 Philipp De Spiegeleer,1 Jan Sermon,1 Kristel Hauben,1 Anne Farewell,2 Thomas Nyström,2 and Chris W. Michiels1*

Laboratory of Food Microbiology, Katholieke Universiteit Leuven, Leuven, Belgium,1 Department of Cell and Molecular Biology-Microbiology, Göteborg University, Göteborg, Sweden2

Received 13 October 2003/ Accepted 11 February 2004

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.

* Corresponding author. Mailing address: Laboratory of Food Microbiology, K. U. Leuven, Kasteelpark Arenberg 22, B-3001 Heverlee, Belgium. Phone: 32-(0)16-32 15 78. Fax: 32-(0)16-32 19 60. E-mail: Chris.Michiels{at}agr.kuleuven.ac.be.

Applied and Environmental Microbiology, May 2004, p. 2660-2666, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.2660-2666.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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