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Abstract
The ompB region on the Escherichia coli chromosome codes for two genes, ompR and envZ, which are required for the osmolarity sensitive biosynthetic regulation of the outer membrane matrix proteins (porins), OmpF and ompC. A part of the ompB region containing the ompR gene has been cloned (Wurtzel, E. T., Movva, N. R., Ross, F. L., and Inouye, M. (1981) J. Mol. Appl. Genet. 1, 61-69). We have determined the DNA sequence, including the promoter and structural regions encompassed in a 1.3-kilobase pair Ava I-Eco RI subfragment. This fragment codes for the entire ompR gene as well as the 5' end of the envZ gene. The ompR gene codes for a protein of 32,489 daltons, consisting of 284 amino acid residues. This was confirmed by identifying the gene product by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and determining a partial amino acid sequence of the NH2-terminal region of the gene product. A sequence of 57 amino acid residues located in the COOH-terminal region of the protein is extremely basic. It contains 10 arginine plus lysine residues in contrast to 1 glutamic acid residue in this region. In vitro transcription of the DNA from this region indicates that ampR and envZ are co-transcribed as a polycistronic mRNA from a promoter located 5' to the ompR gene. Translation of the am pR gene terminates at two tandem TAS codons and translation of the envZ gene initiates 29 nucleotides downstream. Cloning of the promoter region of ompB at a site 5' to the structural portion of the beta-galactosidase gene indicates that transcription of ompB is under positive control by cAMP.
Highlights
The envZ genetogether with the ompR gene is located entidy synthesized in media of high osmolarity (6, 7)
When the envZ gene was subcloned into an expression-cloningvector (PIN-II-A), a protein of an apparent molecular weight of 44,000 was lated such that thetotal amount of these proteins in the outer membrane remains constant
We report the DNA sequence of the enuZ gene and the identification of the envZ gene product by employing an expression-cloning vector developed in our laboratory
Summary
The envZ genetogether with the ompR gene is located entidy synthesized in media of high osmolarity (6, 7). Together with the DNA sequence of the ompR gene determined inthe preceding paper
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