Human immunodeficiency virus type 1 N-terminal capsid mutants that exhibit aberrant core morphology and are blocked in initiation of reverse transcription in infected cells

被引:125
作者
Tang, SX
Murakami, T
Agresta, BE
Campbell, S
Freed, EO
Levin, JG
机构
[1] NICHHD, Mol Genet Lab, NIH, Bethesda, MD 20892 USA
[2] NIAID, Mol Microbiol Lab, Bethesda, MD 20892 USA
[3] NCI, HIV Drug Resistance Program, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA
关键词
D O I
10.1128/JVI.75.19.9357-9366.2001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A group of conserved hydrophobic residues faces the interior of the coiled-coil-like structure within the N-terminal domain of the human immunodeficiency virus type 1 (HIV-1) capsid protein (CA). It has been suggested that these residues are important for maintaining stable structure and functional activity. To investigate this possibility, we constructed two HIV-1 clones, in which Trp23 or Phe40 was changed to Ala. We also constructed a third mutant, D51A, which has a mutation that destroys a salt bridge between Pro1 and Asp51. All three mutants are replication defective but produce virus particles. Mutant virions contain all of the viral proteins, although the amount and stability of CA are decreased and levels of virion-associated integrase are reduced. The mutations do not affect endogenous reverse transcriptase activity; however, the mutants are blocked in their ability to initiate reverse transcription in infected cells and no minus-strand strong-stop DNA is detected. The defect in reverse transcription is associated with striking defects in the morphology of mutant virus cores, as determined by transmission electron microscopy. Our data indicate that the mutations made in this study disrupt CA structure and prevent proper maturation of virus cores. We propose that this results in a defect in core stability or in an early postentry event preceding reverse transcription.
引用
收藏
页码:9357 / 9366
页数:10
相关论文
共 61 条
[1]   Isolation of human immunodeficiency virus type 1 cores:: Retention of Vpr in the absence of p6gag [J].
Accola, MA ;
Öhagen, Å ;
Göttlinger, HG .
JOURNAL OF VIROLOGY, 2000, 74 (13) :6198-6202
[2]   PRODUCTION OF ACQUIRED IMMUNODEFICIENCY SYNDROME-ASSOCIATED RETROVIRUS IN HUMAN AND NONHUMAN CELLS TRANSFECTED WITH AN INFECTIOUS MOLECULAR CLONE [J].
ADACHI, A ;
GENDELMAN, HE ;
KOENIG, S ;
FOLKS, T ;
WILLEY, R ;
RABSON, A ;
MARTIN, MA .
JOURNAL OF VIROLOGY, 1986, 59 (02) :284-291
[3]   Amino acid substitutions in the CA protein of moloney murine leukemia virus that block early events in infection [J].
Alin, K ;
Goff, SP .
VIROLOGY, 1996, 222 (02) :339-351
[4]   Second-site suppressors of Rous sarcoma virus CA mutations: Evidence for interdomain interactions [J].
Bowzard, JB ;
Wills, JW ;
Craven, RC .
JOURNAL OF VIROLOGY, 2001, 75 (15) :6850-6856
[5]   Cyclophilin A is required for an early step in the life cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription [J].
Braaten, D ;
Franke, EK ;
Luban, J .
JOURNAL OF VIROLOGY, 1996, 70 (06) :3551-3560
[6]  
Brown P. O., 1997, P161
[7]   Viral DNA synthesis defects in assembly-competent Rous sarcoma virus CA mutants [J].
Cairns, TM ;
Craven, RC .
JOURNAL OF VIROLOGY, 2001, 75 (01) :242-250
[8]   FUNCTIONAL DOMAINS OF THE CAPSID PROTEIN OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 [J].
DORFMAN, T ;
BUKOVSKY, A ;
OHAGEN, A ;
HOGLUND, S ;
GOTTLINGER, HG .
JOURNAL OF VIROLOGY, 1994, 68 (12) :8180-8187
[9]   HIV-1 nucleocapsid protein induces ''maturation'' of dimeric retroviral RNA in vitro [J].
Feng, YX ;
Copeland, TD ;
Henderson, LE ;
Gorelick, RJ ;
Bosche, WJ ;
Levin, JG ;
Rein, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (15) :7577-7581
[10]   Proline residues in the HIV-1NH2-terminal capsid domain:: Structure determinants for proper core assembly and subsequent steps of early replication [J].
Fitzon, T ;
Leschonsky, B ;
Bieler, K ;
Paulus, C ;
Schröder, J ;
Wolf, H ;
Wagner, R .
VIROLOGY, 2000, 268 (02) :294-307