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PMID: 8474159 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Basic amino acids flanking the zinc finger of Moloney murine leukemia virus nucleocapsid protein NCp10 are critical for virus infectivity.

Journal of virology ·Vol. 67 ·No. 5 ·1993-05-00 ·Pages 2537-45

Housset V, De Rocquigny H, Roques BP, Darlix JL

Abstract

Nucleocapsid (NC) protein NCp10 of Moloney murine leukemia virus is encoded by the 3' domain of gag and contains a zinc finger surrounded by basic amino acids. During virion assembly, NC protein is necessary for core formation and the NC zinc finger is required for the packaging of the genomic RNA dimer. In vitro NCp10 has RNA-binding and -annealing activities critical for virus infectivity, since NCp10 promotes dimerization of viral RNA containing the Psi packaging element and annealing of replication primer tRNA(Pro) to the initiation site of reverse transcription (primer-binding site). To investigate the role of the basic amino acids flanking the NCp10 zinc finger, neutral residues were substituted for the basic amino acids and the effects of these mutations in vivo on virus assembly and infectivity and in vitro on the RNA-annealing activity of NCp10 were analyzed. Here we report that the substitution of 1 or 2 neutral amino acids for the basic residues did not impair the production of mature virions but that infectivity was either moderately or strongly attenuated. When more than 2 basic residues were replaced by neutral amino acids, viruses were poorly infectious because of a severe defect in genomic RNA dimer packaging and initiation of reverse transcription. In vitro NCp10-derived peptides with similar mutations were chemically synthesized and were found to be either fully or partially active or completely inactive. These data indicate that the basic residues flanking the zinc finger of NCp10 are required for the production of infectious Moloney murine leukemia virus virions.

MeSH Terms
3T3 Cells Amino Acid Sequence Amino Acids, Diamino/genetics Animals Base Sequence Gene Products, gag/genetics,metabolism Mice Molecular Sequence Data Moloney murine leukemia virus/genetics,growth & development Mutagenesis, Site-Directed Peptide Fragments/metabolism RNA, Viral/analysis Structure-Activity Relationship Transcription, Genetic Viral Core Proteins/genetics,metabolism Viral Proteins/analysis Virion/chemistry Virus Replication Zinc Fingers
Chemicals
Amino Acids, Diamino Gene Products, gag Peptide Fragments RNA, Viral Viral Core Proteins Viral Proteins retroviral nucleocapsid protein NCp10, Moloney murine leukemia virus
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Housset V
LaboRetro Institut National de la Santé et de la Recherche Médicale, Ecole Normale Supérieure de Lyon, France.
De Rocquigny H
Roques B P
Darlix J L
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.67.5.2537-2545.1993
Published
1993-05-00
Pages
2537-45
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237573
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.67.5.2537-2545.1993 PMC Full Text View journal details
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