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PMID: 8415644 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

RNase E activity is conferred by a single polypeptide: overexpression, purification, and properties of the ams/rne/hmp1 gene product.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 90 ·No. 19 ·1993-10-01 ·Pages 9006-10

Cormack RS, Genereaux JL, Mackie GA

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.

Related Genes
ams hmp1 rne
MeSH Terms
Base Sequence Binding Sites DNA Primers Electrophoresis, Polyacrylamide Gel Endoribonucleases/genetics,isolation & purification,metabolism Escherichia coli/enzymology,genetics Genes, Bacterial Molecular Sequence Data Molecular Weight Plasmids Polymerase Chain Reaction Recombinant Proteins/biosynthesis,isolation & purification,metabolism
Chemicals
DNA Primers Recombinant Proteins Endoribonucleases ribonuclease E
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Cormack R S
Department of Biochemistry, University of Western Ontario, London, Canada.
Genereaux J L
Mackie G A
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24 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.90.19.9006
Published
1993-10-01
Pages
9006-10
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC47490
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.90.19.9006 PMC Full Text View journal details
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