Functional organization of the murine leukemia virus reverse transcriptase: characterization of a bacterially expressed AKR DNA polymerase deficient in RNase H activity.

Journal of virology ·Vol. 62 ·No. 11 ·1988-11-00 ·Pages 4376-80

Levin JG, Crouch RJ, Post K, Hu SC, McKelvin D, Zweig M, Court DL, Gerwin BI

Abstract

The functional organization of the murine leukemia virus reverse transcriptase was investigated by expressing a molecular clone containing AKR MuLV reverse transcriptase-coding sequences in Escherichia coli. A purified preparation of the expressed enzyme (pRT250 reverse transcriptase) consisted primarily of a 69-kilodalton protein that has normal levels of murine leukemia virus polymerase activity but 10-fold-reduced levels of RNase H compared with the viral enzyme. The deficit in RNase H activity was correlated with the absence of 60 to 65 amino acids normally present at the carboxyl end of murine leukemia virus reverse transcriptase. The results provide additional experimental evidence for the localization of polymerase and RNase H domains to the N- and C-terminal regions of reverse transcriptase, respectively.

MeSH Terms

AKR murine leukemia virus/enzymology,genetics Amino Acid Sequence Base Composition Binding Sites Cloning, Molecular Endoribonucleases/metabolism Escherichia coli/genetics Leukemia Virus, Murine/enzymology Molecular Sequence Data RNA-Directed DNA Polymerase/genetics,physiology Ribonuclease H Viral Proteins/genetics,physiology

Chemicals

Viral Proteins RNA-Directed DNA Polymerase Endoribonucleases Ribonuclease H

Authors & Affiliations

8 authors, click to expand affiliations / ORCID

Levin J G

Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.

Crouch R J

Post K

Hu S C

McKelvin D

Zweig M

Court D L

Gerwin B I

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Article Info

Journal

Journal of virology

Abbr.

J Virol

ISSN

0022-538X

DOI

10.1128/JVI.62.11.4376-4380.1988

Published

1988-11-00

Pages

4376-80

Language

English

Region

United States

NLM ID

0113724

PMCID

PMC253878

Subset

External Links

PubMed source DOI: 10.1128/JVI.62.11.4376-4380.1988 PMC Full Text View journal details

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Functional organization of the murine leukemia virus reverse transcriptase: characterization of a bacterially expressed AKR DNA polymerase deficient in RNase H activity.

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