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Journal of Virology, March 2001, p. 2616-2626, Vol. 75, No. 6
Integrated Program in Cellular, Molecular,
and Biophysical Studies,1 Department of
Biochemistry and Molecular Biophysics,2
Department of Microbiology,3 and
Howard Hughes Medical Institute,4
Columbia University College of Physicians and Surgeons, New York, New
York 10032
Received 26 September 2000/Accepted 19 December 2000
The nucleocapsid (NC) protein of retroviruses is a small nucleic
acid-binding protein important in virion assembly and in the
encapsidation of the viral RNA genome into the virion particle. Multiple single-amino-acid substitutions were introduced into the NC of
Moloney murine leukemia virus to examine further its role in viral
replication. Two residues were shown to play important roles in the
early events of replication. Unlike viruses with previously
characterized NC mutations, these viruses showed no impairment in the
late events of replication. Viruses containing the substitutions L21A
and K30A expressed the normal complement of properly processed viral
Gag proteins. Analysis of the RNA content of mutant virions revealed
normal levels of unspliced and spliced viral RNA, and the
tRNAPro primer was properly annealed to the primer binding
site on the viral genome. The virions demonstrated no defect in
initiation of reverse transcription using the endogenous tRNA primer or
in the synthesis of long viral DNA products in vitro. Nonetheless, viruses possessing these NC mutations demonstrated significant defects
in the synthesis and accumulation of viral DNA products in vivo.
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.6.2616-2626.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Identification of Residues of the Moloney Murine Leukemia
Virus Nucleocapsid Critical for Viral DNA Synthesis In
Vivo
*
Corresponding author. Mailing address: Department of
Biochemistry and Molecular Biophysics, Columbia University College of Physicians and Surgeons, New York, NY 10032. Phone: (212) 305-3794. Fax: (212) 305-8692. E-mail:
goff{at}cuccfa.ccc.columbia.edu.
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