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Nucleic Acids Research, 1985, Vol. 13, No. 13 4677-4685
© 1985

Articles

The DNA sequence of the gene (rnc) encoding ribonuclease III of Escherichia coli

Paul E. March, Joohong Ahnn and Masayori Inouye

Department of Biochemistry, State University of New York Stony Brook, NY 11794, USA

Received May 10, 1985. Accepted June 5, 1985.

The DNA sequence of a 1,076 base pair Bg1I-BamHI fragment containing the entire rnc gene for ribonuclease III (RNase III) was determined. An open reading frame of 681 base pairs was found in this region which encodes a protein of 227 amino acid residues (calculated molecular weight = 25,218).

When this open reading frame was cloned into a high expression vector, pIN-III, a protein of apparent molecular weight of 26,000 was produced upon induction of the cloned gene. This product accounted for up to 5% of the total cellular protein, and comigrated with purified RNase III. RNase III enzyme activity was induced in parallel with the production of the 26,000 molecular weight protein. A putative promoter was found 170 base pairs upstream from the initiation codon. In the long leader region a very stable stem-bulge-stem structure was found which closely resembles typical RNase III cleavage sites. This structure may be cleaved by RNase III to auto-regulate the expression of the rnc gene.


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