Clustered organization and transcriptional analysis of a family of five csp genes of Lactococcus lactis MG1363.
A family of genes encoding cold-shock proteins, named cspA, cspB, cspC, cspD and cspE, was cloned and sequenced from Lactococcus lactis MG1363. The genes cspA and cspB and the genes cspC and cspD are located in tandem repeats, an organization of csp genes that has never been encountered before. The five genes encode small (7.1-7.6 kDa) proteins with high mutual sequence identities (up to 85%) and high identities (about 45-65%) with the major cold-shock proteins from Escherichia coli (CspA) and Bacillus subtilis (CspB). Northern-blot analysis revealed single transcripts of about 300 nucleotides for each csp gene and showed that cspA, cspB, cspC and cspD mRNA levels were strongly increased upon cold shock to 10 degrees C (about 10-, 40-, 10- and 30-fold compared to 30 degrees C, respectively), whereas the cspE mRNA level was not increased. The expression of the cold-induced csp genes was highest in the 6-8 h lag phase after cold shock. A differential expression in time, in which cspA and cspC were maximally expressed at 2 h and cspB and cspD at 4 h after cold shock, was observed. The -35 and -10 regions of the five promoters were identified and transcriptional start sites were mapped in each case by primer extension at different temperatures which confirmed that regulation takes place at the transcriptional level. Significant differences were observed between the 5'-untranslated leader regions of the four cold-induced csp genes and the corresponding region of the non-cold-induced cspE gene.
Woufers
Jeroen A
JA
Laboratory for Food Microbiology, Food Science Group, Wageningen University and Research CenterWageningenThe Netherlands.|Microbial Ingredients Section, NlZO Food ResearchPO Box 20, 6710 BA EdeThe Netherlands.
Sander
Jan-Willem
JW
Department of Genetics, Groningen UniversityHarenThe Netherlands.
Kok
Jan
J
Department of Genetics, Groningen UniversityHarenThe Netherlands.
de Vos
Willem M
WM
Microbial Ingredients Section, NlZO Food ResearchPO Box 20, 6710 BA EdeThe Netherlands.
Kuipers
Oscar P
OP
Microbial Ingredients Section, NlZO Food ResearchPO Box 20, 6710 BA EdeThe Netherlands.
Abee
Tjakko
T
Laboratory for Food Microbiology, Food Science Group, Wageningen University and Research CenterWageningenThe Netherlands.
D020121:5' Untranslated Regions / Q000235:genetics; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D003080:Cold Temperature; D016384:Consensus Sequence / Q000235:genetics; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics; D013294:Lactococcus lactis / Q000235:genetics* / Q000254:growth & development; D008969:Molecular Sequence Data; D005810:Multigene Family; D016366:Open Reading Frames / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D016601:RNA-Binding Proteins; D016415:Sequence Alignment
D020121:5' Untranslated Regions; D001426:Bacterial Proteins; D002352:Carrier Proteins; C491179:CspB protein, E coli; C088099:CspC protein, bacteria; C088098:CspD protein, bacteria; C088097:CspE protein, E coli; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D016601:RNA-Binding Proteins; C062260:cold shock protein CS7.4, Bacteria; C077188:cold-shock protein CspB, Bacteria; C490241:cspC protein, E coli; C490240:cspH protein, E coli