TY - JOUR
T1 - The DNA sequence of the gene (rnc) encoding ribonuclease III of Escherichia coli
AU - March, Paul E.
AU - Ahnn, Joohong
AU - Inouye, Masayori
N1 - Funding Information:
ACKNOWLEDGEMENTS We thank Dr. H. Robertson for assaying RNase III activity in a cell lysate and for purified RNase III preparation used as a standard in SDS-polyacrylamide gel electrophoresis. We are grateful to Jack Coleman for critical reading of the manuscript. We thank the National Institute of General Medical Sciences (Grant GM19043) and the American Cancer Society (Grant NP387I) for support of this research. P.E.M. is an American Cancer Society postdoctoral fellow.
PY - 1985/7/11
Y1 - 1985/7/11
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0022423457&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0022423457&partnerID=8YFLogxK
U2 - 10.1093/nar/13.13.4677
DO - 10.1093/nar/13.13.4677
M3 - Article
C2 - 3895158
AN - SCOPUS:0022423457
SN - 0305-1048
VL - 13
SP - 4677
EP - 4685
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 13
ER -