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Journal of Virology, August 2000, p. 7250-7260, Vol. 74, No. 16
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Infectivity of Moloney Murine Leukemia Virus Defective in Late
Assembly Events Is Restored by Late Assembly Domains of Other
Retroviruses
Bing
Yuan,1
Stephen
Campbell,2
Eran
Bacharach,3
Alan
Rein,2 and
Stephen P.
Goff3,4,*
Integrated Program in Cellular, Molecular and
Biophysical Studies,1 Howard Hughes
Medical Institute,4 and Department of
Biochemistry and Molecular Biophysics,3 College
of Physicians and Surgeons, Columbia University, New York, New York
10032, and HIV Drug Resistance Program, National Cancer
Institute
Frederick Cancer Research and Development Center,
Frederick, Maryland 217022
Received 3 March 2000/Accepted 12 May 2000
The p12 region of the Moloney murine leukemia virus (M-MuLV) Gag
protein contains a PPPY motif important for efficient virion assembly and release. To probe the function of the PPPY motif, a series
of insertions of homologous and heterologous motifs from other
retroviruses were introduced at various positions in a mutant gag gene lacking the PPPY motif. The assembly defects of
the PPPY deletion mutant could be rescued by insertion of a wild-type
PPPY motif and flanking sequences at several ectopic positions in the Gag protein. The late assembly domain (L-domain) of Rous sarcoma virus
(RSV) or human immunodeficiency virus type 1 (HIV-1) could also fully
or partially restore M-MuLV assembly when introduced into matrix, p12,
or nucleocapsid domains of the mutant M-MuLV Gag protein lacking the
PPPY motif. Strikingly, mutant viruses carrying the RSV or the HIV-1
L-domain at the original location of the deleted PPPY motif were
replication competent in rodent cells. These data suggest that the PPPY
motif of M-MuLV acts in a partially position-independent manner and is
functionally interchangeable with L-domains of other retroviruses.
Electron microscopy studies revealed that deletion of the entire
p12 region resulted in the formation of tube-like rather than spherical
particles. Remarkably, the PPPY deletion mutant formed chain structures
composed of multiple viral particles linked on the cell surface. Many
of the mutants with heterologous L-domains released virions with
wild-type morphology.
*
Corresponding author. Mailing address: Department of
Biochemistry and Molecular Biophysics, Howard Hughes Medical Institute, Columbia University College of Physicians and Surgeons, 701 W. 168th
St., New York, NY 10032. Phone: (212) 305-3794. Fax: (212) 305-8692. E-mail: goff{at}cuccfa.ccc.columbia.edu.
Journal of Virology, August 2000, p. 7250-7260, Vol. 74, No. 16
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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