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Identification of the promoter region of the Escherichia coli major cold shock gene, cspA.

Department of Biochemistry, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey at Rutgers, Piscataway 08854-5635.

ABSTRACT

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.

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