Tanabe H, Goldstein J, Yang M, Inouye M
Department of Biochemistry, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey at Rutgers, Piscataway 08854-5635.
J Bacteriol. 1992 Jun;174(12):3867-73. doi: 10.1128/jb.174.12.3867-3873.1992.
The major cold shock protein of Escherichia coli, CS7.4, is produced at a level of 13% of total protein synthesis upon a temperature shift from 37 to 10 degrees C. The transcription of its gene (cspA) was found to be tightly regulated and induced only at low temperature. In addition, the cspA mRNA was extremely unstable at 37 degrees C, so that CS7.4 production was hardly detected when the culture temperature was shifted from 15 degrees C to 37 degrees C. The transcription initiation site (+1) was identified. In vivo footprinting demonstrated that the region from bases -35 to -73 was protected from chemical modification, and gel mobility shift analysis showed that a cold-shocked cell extract contained a factor(s) specifically bound to the fragment containing the sequence between bases -63 and -92. This factor was synthesized de novo only at low temperature, and its synthesis was inhibited by chloramphenicol. Possible functions of this factor are discussed.
大肠杆菌的主要冷休克蛋白CS7.4,在温度从37℃转变为10℃时,其产生量占总蛋白合成量的13%。发现其基因(cspA)的转录受到严格调控,且仅在低温下被诱导。此外,cspA mRNA在37℃时极其不稳定,因此当培养温度从15℃转变为37℃时,几乎检测不到CS7.4的产生。确定了转录起始位点(+1)。体内足迹分析表明,从碱基-35到-73的区域受到保护不被化学修饰,凝胶迁移率变动分析显示,冷休克细胞提取物含有一种与包含碱基-63到-92之间序列的片段特异性结合的因子。这种因子仅在低温下重新合成,其合成受到氯霉素的抑制。讨论了该因子可能的功能。