Cloning and characterization of a gene encoding a cold-shock protein in Lactobacillus casei
Nicolas Sauvageot,
Sophie Beaufils,
Alain Mazé,
Josef Deutscher,
Axel Hartke,
Nicolas Sauvageot
Laboratoire de Microbiologie de l'Environnement, Université de Caen, Esplanade de la Paix, Caen Cedex, France
Search for more papers by this authorSophie Beaufils
Laboratoire de Microbiologie de l'Environnement, Université de Caen, Esplanade de la Paix, Caen Cedex, France
Search for more papers by this authorAlain Mazé
Laboratoire de Microbiologie et Génétique Moléculaire, Thiverval-Grignon, France
Search for more papers by this authorJosef Deutscher
Laboratoire de Microbiologie et Génétique Moléculaire, Thiverval-Grignon, France
Search for more papers by this authorAxel Hartke
Laboratoire de Microbiologie de l'Environnement, Université de Caen, Esplanade de la Paix, Caen Cedex, France
Search for more papers by this authorNicolas Sauvageot,
Sophie Beaufils,
Alain Mazé,
Josef Deutscher,
Axel Hartke,
Nicolas Sauvageot
Laboratoire de Microbiologie de l'Environnement, Université de Caen, Esplanade de la Paix, Caen Cedex, France
Search for more papers by this authorSophie Beaufils
Laboratoire de Microbiologie de l'Environnement, Université de Caen, Esplanade de la Paix, Caen Cedex, France
Search for more papers by this authorAlain Mazé
Laboratoire de Microbiologie et Génétique Moléculaire, Thiverval-Grignon, France
Search for more papers by this authorJosef Deutscher
Laboratoire de Microbiologie et Génétique Moléculaire, Thiverval-Grignon, France
Search for more papers by this authorAxel Hartke
Laboratoire de Microbiologie de l'Environnement, Université de Caen, Esplanade de la Paix, Caen Cedex, France
Search for more papers by this author Axel Hartke, Laboratoire de Microbiologie de l'Environnement (EA956 USC I.N.R.A. 2017), IBFA, Université de Caen, Esplanade de la Paix, F-14032 Caen Cedex, France. Tel.: +33 2 31 56 54 04; fax: +33 2 31 56 53 11; e-mail: [email protected]
Editor: Robert Burne
Abstract
One csp-like gene, called cspA, from the lactic acid bacterium Lactobacillus casei was identified by an inverse polymerase chain reaction approach based on degenerate primers. cspA encodes a protein of 66 amino acid residues, which displays at least 74% identity with Csp proteins of Lactobacillus genera. Northern blot analysis revealed that cspA is transcribed monocistronically and that its expression is induced after a temperature downshift from 37°C to 20°C. The transcriptional start site has been determined and is situated 98 bp upstream of the initiation codon. A cspA mutant strain was constructed and it showed reduced growth rate compared with the wild type at both optimal and low temperatures, demonstrating that CspA plays an important role in the physiology of L. casei.
References
- Acedo-Felix E & Perez-Martinez G (2003) Significant differences between Lactobacilluscasei subsp. casei ATCC 393 T and a commonly used plasmid-cured derivative revealed by a polyphasic study. Int J Syst Evol Microbiol 53: 67 – 75.
- Altschul SF, Gish W, Myers EW & Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215: 403 – 410.
- Bae W, Jones PG & Inouye M (1997) CspA, the major cold shock protein of Escherichiacoli, negatively regulates its own gene expression. J Bacteriol 179: 7081 – 7088.
- Bae W, Xia B, Inouye M & Severinov K (2000) Escherichia coli CspA-family RNA chaperones are transcription antiterminators. Proc Natl Acad Sci USA 97: 7784 – 7789.
- Berthier F, Zagorec M, Champomier-Vergès M, Ehrlich SD & Morel-Deville F (1996) Efficient transformation of Lactobacillus sake by electroporation. Microbiology 142: 1273 – 1279.
- Bolotin A, Wincker P, Mauger S, Jaillon O, Malarme K, Weissenbach J, Ehrlich SD & Sorokin A (2001) The complete genome sequence of the lactic acid bacterium Lactococcuslactis ssp. lactis IL1403. Genome Res 11: 731 – 753.
- Brandi A, Pon CL & Gualerzi CO (1994) Interaction of the main cold shock protein CS7.4 (CspA) of Escherichia coli with the promoter region of hns. Biochimie 76: 1090 – 1098.
- Brandi A, Pietroni P, Gualerzi CO & Pon CL (1996) Post-transcriptional regulation of CspA expression in Escherichiacoli. Mol Microbiol 19: 231 – 240.
- Chapot-Chartier MP, Schouler C, Lepeuple AS, Gripon JC & Chopin MC (1997) Characterization of cspB, a cold-shock-inducible gene from Lactococcuslactis, and evidence for a family of genes homologous to the Escherichia coli cspA major cold shock gene. J Bacteriol 179: 5589 – 5593.
- Connil N, Le Breton Y, Dousset X, Auffray Y, Rincé A & Prévost H (2002) Identification of the Enterococcusfaecalis tyrosine decarboxylase operon involved in tyramine production. Appl Environ Microbiol 68: 3537 – 3544.
- Darbon E, Servant P, Poncet S & Deutscher J (2002) Antitermination by GlpP, catabolite repression via CcpA and inducer exclusion triggered by P-GlpK dephosphorylation control Bacillus subtilisglpFK expression. Mol Microbiol 43: 1039 – 1052.
- Derzelle S, Hallet B, Francis KP, Ferain T, Delcour J & Hols P (2000) Changes in cspL, cspP, and cspC mRNA abundance as a function of cold shock and growth phase in Lactobacillus plantarum. J Bacteriol 182: 5105 – 5113.
- Dufour A, Rincé A, Uguen P & Le Pennec JP (2000) IS1675, a novel lactococcal insertion element, forms a transposon-like structure including the lacticin 481 lantibiotic operon. J Bacteriol 182: 5600 – 5605.
- Fang L, Jiang W, Bae W & Inouye M (1997) Promoter independent cold-shock induction of cspA and its derepression at 37°C by mRNA stabilization. Mol Microbiol 23: 355 – 364.
- Fang L, Xia B & Inouye M (1999) Transcription of CspA, the gene for the major cold-shock protein of Escherichia coli, is negatively regulated at 37 degrees C by the 5′-untranslated region of its mRNA. FEMS Microbiol Lett 176: 39 – 43.
- Francis KP & Stewart GS (1997) Detection and speciation of bacteria through PCR using universal major cold-shock protein primer oligomers. J Ind Microbiol Biotechnol 19: 286 – 293.
- Graumann P & Marahiel MA (1996) Some like it cold: response of microorganisms to cold shock. Arch Microbiol 166: 293 – 300.
- Graumann P & Marahiel MA (1999) Cold shock proteins CspB and CspC are major stationary-phase-induced proteins in Bacillus subtilis. Arch Microbiol 166: 293 – 300.
- Graumann P, Schroder K, Schmid R & Marahiel MA (1996) Cold shock stress-induced proteins in Bacillus subtilis. J Bacteriol 178: 4611 – 4619.
- Graumann P, Wendrich TM, Weber MHW, Schröder K & Marahiel MA (1997) A family of cold shock proteins in Bacillus subtilis is essential for cellular growth and for efficient protein synthesis at optimal and low temperatures. Mol Microbiol 25: 741 – 756.
- van de Guchte M, Serror P, Chervaux C, Smokvina T, Ehrlich SD & Maguin E (2002) Stress response in lactic acid bacteria. Antonie van Leeuwenhoek 82: 187 – 216.
- Jiang W, Hou Y & Inouye M (1997) CspA, the major cold-shock protein of Escherichia coli, is an RNA chaperone. J Biol Chem 272: 196 – 202.
- Jones PG & Inouye M (1994) The cold-shock response a hot topic. Mol Microbiol 11: 811 – 818.
- Jones PG, Mitta M, Kim Y, Jiang W & Inouye M (1996) Cold shock induces a major ribosomal-associated protein that unwinds double stranded RNA in Escherichia coli. Proc Natl Acad Sci USA 93: 76 – 80.
- La Teana A, Brandi A, Falconi M, Spurio R, Pon CL & Gualerzi CO (1991) Identification of a cold shock transcriptional enhancer of the Escherichiacoli gene encoding nucleoid protein H-NS. Proc Natl Acad Sci USA 88: 10907 – 10911.
- Landsman D (1992) RNP-1, an RNA-binding motif is conserved in the DNA-binding cold shock domain. Nucleic Acids Res 20: 2861 – 2864.
- Lee SJ, Xie A, Jiang W, Etchegaray JP, Jones PG & Inouye M (1994) Family of the major cold-shock protein, CspA (CS7.4), of Escherichia coli, whose members show a high sequence similarity with the eukaryotic Y-box bindingproteins. Mol Microbiol 11: 833 – 839.
- Leloup L, Ehrlich SD, Zagorec M & Morel-Deville F (1997) Single-crossover integration in the Lactobacillus sake chromosome and insertional inactivation of the ptsI and lacL genes. Appl Environ Microbiol 63: 2117 – 2123.
- De Man JC, Rogosa M & Sharp ME (1960) A medium for the cultivation of Lactobacilli. J Appl Bacteriol 23: 130 – 135.
- Mayo B, Derzelle S, Fernández M, Léonard C, Ferain T, Hols P, Suárez JE & Delcour J (1997) Cloning and characterization of cspL and cspP, two cold-inducible genes from Lactobacillus plantarum. J Bacteriol 179: 3039 – 3042.
- Moll I, Huber M, Grill S, Sairafi P, Mueller F, Brimacombe R, Londei P & Blasi U (2001) Evidence against an interaction between the mRNA downstream box and 16S rRNA intranslation initiation. J Bacteriol 183: 3499 – 3505.
- Phadtare S, Yamanaka K & Inouye M (2000) The cold shock response. Bacterial Stress Response ( G Strorz & R Hengge-Aronis, eds), pp. 33 – 45. ASM Press, Washington, DC.
-
Sambrook J,
Fritsch EF &
Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
10.1111/j.1095-8312.1996.tb01434.x Google Scholar
- Schroder K, Graumann P, Schnuchel A, Holak TA & Marahiel MA (1995) Mutational analysis of the putative nucleic acid-binding surface of the cold-shock domain, CspB, revealed an essential role of aromatic and basic residues in binding of single-stranded DNA containing the Y-box motif. Mol Microbiol 16: 699 – 708.
- Serror P, Dervyn R, Ehrlich SD & Maguin E (2003) Csp-like genes of Lactobacillus delbrueckii ssp. Bulgaricus and their response to cold shock. FEMS Microbiol Lett 226: 323 – 330.
- Wang N, Yamanaka K & Inouye M (1999) CspI, the ninth member of the CspA family of Escherichia coli, is induced upon cold shock. J Bacteriol 181: 1603 – 1609.
- Weber MH, Beckering CL & Marahiel MA (2001) Complementation of cold shock proteins by translation initiation factor IF1 in vivo. J Bacteriol 183: 7381 – 7386.
- Wouters JA, Jeynov B, Rombouts FM, de Vos WM, Kuipers OP & Abee T (1999) Analysis of the role of 7 kDa cold-shock proteins of Lactococcus lactis MG1363 in cryoprotection. Microbiology 145: 3185 – 3194.
- Wouters JA, Frenkiel H, De Vos WM, Kuipers OP & Abee T (2001a) Cold shock proteins of Lactococcus lactis MG1363 are involved in cryoprotection and in the production of cold-induced proteins. Appl Environ Microbiol 67: 5171 – 5178.
- Wouters JA, Frenkiel H, de Vos WM, Kuipers OP & Abee T (2001b) Cold shock proteins of Lactococcus lactis MG1363 are involved in cryoprotection and in the production of cold-induced proteins. Appl Environ Microbiol 67: 5171 – 5178.
- Xia B, Ke H & Inouye M (2001) Acquirement of cold sensitivity by quadruple deletion of the cspA family and its suppression by PNPase S1 domain in Escherichiacoli. Mol Microbiol 40: 179 – 188.
- Yamanaka K, Fang L & Inouye M (1998) The CspA family in Escherichia coli: multiple gene duplication for stress adaptation. Mol Microbiol 27: 247 – 255.
- Yamanaka K, Mitta M & Inouye M (1999) Mutation analysis of the 5′ untranslated region of the cold shock CspA mRNA of Escherichiacoli. J Bacteriol 181: 6284 – 6291.
