A Mutation Affecting the Regulation of a secA-lacZ Fusion Defines a New sec Gene

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A Mutation Affecting the Regulation of a secA-lacZ Fusion Defines a New sec Gene

P. D. Riggs, A. I. Derman and J. Beckwith
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115

It was shown previously that the secA gene of Escherichia coli is derepressed in cells that have a defect in protein export. Here it is demonstrated that the {beta}-galactosidase produced by a secA-lacZ gene fusion strain is regulated in the same way. Studies on the fusion strain reveal that the promoter or a site involved in regulation of the secA gene is located considerably upstream from the structural gene. The properties of the fusion strain provide a new selection for mutants that are defective in protein export. Selection for increased lac expression of a secA-lacZ fusion strain yields mutations in three of the known sec genes, secA, secD and prlA/secY. In addition, mutations in several genes not previously known to affect secA expression were obtained. A mutation in one of these genes causes a pleiotropic defect in protein export and a cold-sensitive growth defect; this gene, which maps at approximately 90 min on the bacterial chromosome, has been named secE.

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				A. Guigueno, J. Dassa, P. Belin, and P. L. Boquet Oversynthesis of a New Escherichia coli Small RNA Suppresses Export Toxicity of DsbA'-PhoA Unfoldable Periplasmic Proteins J. Bacteriol., February 15, 2001; 183(4): 1147 - 1158. [Abstract] [Full Text]

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				Y.-B. Yang, N. Yu, and P. C. Tai SecE-depleted Membranes of Escherichia coli Are Active. SecE IS NOT OBLIGATORILY REQUIRED FOR THE IN VITRO TRANSLOCATION OF CERTAIN PROTEIN PRECURSORS J. Biol. Chem., May 23, 1997; 272(21): 13660 - 13665. [Abstract] [Full Text] [PDF]

				T. Shimoike, T. Taura, A. Kihara, T. Yoshihisa, Y. Akiyama, K. Cannon, and K. Ito Product of a New Gene, syd, Functionally Interacts with SecY when Overproduced in Escherichia coli J. Biol. Chem., March 10, 1995; 270(10): 5519 - 5526. [Abstract] [Full Text] [PDF]

				M. Poritz, H. Bernstein, K Strub, D Zopf, H Wilhelm, and P Walter An E. coli ribonucleoprotein containing 4.5S RNA resembles mammalian signal recognition particle Science, November 23, 1990; 250(4984): 1111 - 1117. [Abstract] [PDF]

				P J Schatz, P D Riggs, A Jacq, M J Fath, and J Beckwith The secE gene encodes an integral membrane protein required for protein export in Escherichia coli. Genes & Dev., July 1, 1989; 3(7): 1035 - 1044. [Abstract] [PDF]

				J Stader, L J Gansheroff, and T J Silhavy New suppressors of signal-sequence mutations, prlG, are linked tightly to the secE gene of Escherichia coli. Genes & Dev., July 1, 1989; 3(7): 1045 - 1052. [Abstract] [PDF]

				M. O. Schmidt, R. M. Brosh Jr., and D. B. Oliver Escherichia coli SecA Helicase Activity Is Not Required in Vivo for Efficient Protein Translocation or Autogenous Regulation J. Biol. Chem., September 28, 2001; 276(40): 37076 - 37085. [Abstract] [Full Text] [PDF]