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In Vitro Properties of RpoS (^S) Mutants of Escherichia coli with Postulated N-Terminal Subregion 1.1 or C-Terminal Region 4 Deleted

J. Gowrishankar,^1* Kaneyoshi Yamamoto,^2, P. R. Subbarayan,^2, and Akira Ishihama^2,

Laboratory of Bacterial Genetics, Centre for DNA Fingerprinting and Diagnostics, Hyderabad 500 076, India,¹ Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan²

Received 25 November 2002/ Accepted 5 February 2003

Derivatives of the stationary-phase sigma factor ^S of Escherichia coli lacking either of two conserved domains, the postulated N-terminal subregion 1.1 or the C-terminal region 4, were shown to be competent in vitro for transcription initiation from several ^S-dependent promoters on supercoiled DNA templates. Unlike wild-type ^S, however, the deletion derivatives were inactive on relaxed templates. The anomalous slow electrophoretic mobility of ^S on denaturing gels was corrected by deletion of subregion 1.1, suggesting that this domain in ^S may be structurally and functionally analogous to subregion 1.1 of ⁷⁰, substitutions in which have previously been shown to rectify the anomalous electrophoretic migration of ⁷⁰ (V. Gopal and D. Chatterji, Eur. J. Biochem. 244:614-618, 1997).

Present address: Department of Agricultural Chemistry, Kinki University, 3327-204 Nakamachi, Nara 631-8505, Japan.

Present address: Department of Biochemistry and Molecular Biology (R-629), University of Miami, School of Medicine, Miami, FL 33101-6129.

Present address: Nippon Institute for Biological Science, Ome, Tokyo 198-0024, Japan.

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