PMID:8415644
| Citation |
Cormack, RS, Genereaux, JL and Mackie, GA (1993) RNase E activity is conferred by a single polypeptide: overexpression, purification, and properties of the ams/rne/hmp1 gene product. Proc. Natl. Acad. Sci. U.S.A. 90:9006-10 |
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| Abstract |
Ribonuclease E, an enzyme that processes pre-5S rRNA from its precursor, is now believed to be the major endoribonuclease participating in mRNA turnover in Escherichia coli. The product of the ams/rne/hmp1 gene, which is required for RNase E activity, was overexpressed, purified to near homogeneity by electroelution from an SDS/polyacrylamide gel, and renatured. The purified polypeptide possesses nucleolytic activity in vitro with a specificity identical to that observed for crude RNase E preparations. In addition, both UV crosslinking and RNA-protein blotting unambiguously showed that the Ams/Rne/Hmp1 polypeptide has a high affinity for RNA. Our results demonstrate that RNase E activity is directly attributable to, and is an inherent property of, an RNA-binding protein, the ams/rne/hmp1 gene product. |
| Links | |
| Keywords |
Base Sequence; Binding Sites; DNA Primers; Electrophoresis, Polyacrylamide Gel; Endoribonucleases/genetics; Endoribonucleases/isolation & purification; Endoribonucleases/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Genes, Bacterial; Molecular Sequence Data; Molecular Weight; Plasmids; Polymerase Chain Reaction; Recombinant Proteins/biosynthesis; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism |
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