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The Enterococcus faecalis gene encoding the novel general stress protein Gsp62

Laboratoire de Microbiologie de l’Environnement, IRBA, Université de Caen, 14032 Caen Cedex, France¹
Laboratoire de Biologie et Chimie Moléculaires, EA 2594, Université de Bretagne Sud, 56000 Vannes, France²

Author for correspondence: Alain Rincé. Tel: +33 2 31 56 55 23. Fax: +33 2 31 56 53 11. e-mail: rince{at}ibba.unicaen.fr

The Enterococcus faecalis general stress protein Gsp62 was purified using two-dimensional gel electrophoresis and its 25 N-terminal amino acid sequence determined. Analysis of the corresponding gene revealed that the gsp62 product is a 172 aa protein. Transcriptional analysis of gsp62 gave evidence for a monocistronic mRNA, the synthesis of which was induced at the onset of stationary phase and in response to heat shock, acid pH, detergents (i.e. SDS or bile salts), ethanol, tert-butyl hydroperoxide, sodium chloride and, to a lesser extent, hydrogen peroxide. 5' rapid amplification of cDNA ends by PCR experiments showed that gsp62 transcription initiates 30 nt upstream of the ATG start codon. Although gsp62 expression was induced in response to various stresses, its disruption had no significant effect on the cell survival after each individual stress. Two-dimensional protein gels from wild-type and mutant cells revealed no pleiotropic effect of the mutation on protein synthesis. Transcriptional fusions with the lacL lacM ß-galactosidase genes showed that an inverted repeat located upstream of the promoter is required for transcriptional induction by environmental stresses but not by entrance into stationary phase. Two distinct mechanisms responding to different signals are thus involved in gsp62 induction.

Keywords: Enterococcus faecalis, general stress protein, Gsp

Abbreviations: tBOOH, tert-butyl hydroperoxide; 2-D, two-dimensional; IR, inverted repeat; RACE, rapid amplification of cDNA ends

^a Present address: Laboratoire de Biologie Cellulaire et Moléculaire, Université du Littoral – Côte d’Opale, Bassin Napoléon, BP120, 62327 Boulogne sur mer, Cedex, France.

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