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Journal of Bacteriology, April 2003, p. 2673-2679, Vol. 185, No. 8
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.8.2673-2679.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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

J. Gowrishankar,1* Kaneyoshi Yamamoto,2,{dagger} P. R. Subbarayan,2,{ddagger} and Akira Ishihama2,§

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 {sigma}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 {sigma}S-dependent promoters on supercoiled DNA templates. Unlike wild-type {sigma}S, however, the deletion derivatives were inactive on relaxed templates. The anomalous slow electrophoretic mobility of {sigma}S on denaturing gels was corrected by deletion of subregion 1.1, suggesting that this domain in {sigma}S may be structurally and functionally analogous to subregion 1.1 of {sigma}70, substitutions in which have previously been shown to rectify the anomalous electrophoretic migration of {sigma}70 (V. Gopal and D. Chatterji, Eur. J. Biochem. 244:614-618, 1997).

* Corresponding author. Mailing address: Laboratory of Bacterial Genetics, Centre for DNA Fingerprinting and Diagnostics, ECIL Rd., Nacharam, Hyderabad 500 076, India. Phone: 91-40-27155609. Fax: 91-40-27155610. E-mail: shankar{at}cdfd.org.in.

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

{ddagger} 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.

Journal of Bacteriology, April 2003, p. 2673-2679, Vol. 185, No. 8
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.8.2673-2679.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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