Chanda Palas Kumar, Mondal Rajkrishna, Sau Keya, Sau Subrata
Department of Biochemistry, Bose Institute, P1/12 - CIT Scheme VII M, Kolkata, W.B., India.
J Basic Microbiol. 2009 Apr;49(2):205-11. doi: 10.1002/jobm.200800065.
Proteins expressed by the bacterial cold shock genes are highly conserved at sequence level and perform various biological functions in both the cold-stressed and normal cells. To study the effects of various agents on the cold shock genes of Staphylococcus aureus, we have cloned the upstream region of cspC from S. aureus Newman and found that the above region possesses appreciable promoter (P(c)) activity even at 37 degrees C. A reporter S. aureus strain CHANDA2, constructed by inserting the P(c)-lacZ transcriptional fusion into S. aureus RN4220 genome, was found to express very low level of beta-galactosidase after cold shock, indicating that low temperature induces P(c) very weakly. Interestingly, transcription from P(c ) was induced very strongly by several antibiotics, hydrogen peroxide and arsenate salt. Cold shock proteins expressed by S. aureus are highly identical at sequence level and bear single-strand nucleic acid binding motifs. A 16 nt downstream box and a 13 nt upstream box were identified at the downstream of initiation codon and at the upstream of ribosome binding site of csp transcripts. Their roles in S. aureus cold shock gene expression have been discussed elaborately.
细菌冷休克基因表达的蛋白质在序列水平上高度保守,在冷应激细胞和正常细胞中均发挥多种生物学功能。为了研究各种试剂对金黄色葡萄球菌冷休克基因的影响,我们克隆了金黄色葡萄球菌纽曼株cspC的上游区域,发现上述区域即使在37℃时也具有明显的启动子(P(c))活性。通过将P(c)-lacZ转录融合体插入金黄色葡萄球菌RN4220基因组构建的报告菌株CHANDA2,在冷休克后表达的β-半乳糖苷酶水平非常低,表明低温对P(c)的诱导作用非常弱。有趣的是,几种抗生素、过氧化氢和砷酸盐能强烈诱导P(c)的转录。金黄色葡萄球菌表达的冷休克蛋白在序列水平上高度相同,并具有单链核酸结合基序。在csp转录本起始密码子下游和核糖体结合位点上游分别鉴定出一个16 nt的下游框和一个13 nt的上游框。文中详细讨论了它们在金黄色葡萄球菌冷休克基因表达中的作用。