Deoxyribonucleoside triphosphate pool imbalances in vivo are associated with an increased retroviral mutation rate

被引:74
作者
Julias, JG [1 ]
Pathak, VK [1 ]
机构
[1] W Virginia Univ, Dept Biochem, Mary Babb Randolph Canc Ctr, Morgantown, WV 26506 USA
关键词
D O I
10.1128/JVI.72.10.7941-7949.1998
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Deoxyribonucleoside triphosphate (dNTP) pool imbalances are associated with an increase in the rate of misincorporation and hypermutation during in vitro reverse transcription reactions. However, the effects of in vivo dNTP pool imbalances on the accuracy of reverse transcription are unknown. We sought to determine the effects of in vivo dNTP pool imbalances on retroviral mutation rates and to test our hypothesis that 3'-azido-3'-deoxythymidine (AZT) increases the retroviral mutation rates through induction of dNTP pool imbalances. D17 cells were treated with thymidine, hydroxyurea (HU), or AZT, and the effects on in vivo dNTP pools were measured. Thymidine and HU treatments induced significant dNTP pool imbalances. In contrast, AZT treatment had very little effect on the dNTP pools. The effects of in vivo dNTP pool imbalances induced by thymidine and HU treatments on the retroviral mutation rates were also determined. Spleen necrosis virus (SNV)-based and murine leukemia virus (MLV)-based retroviral vectors that expressed the lacZ mutant reporter gene were used. The frequencies of inactivating mutations introduced in the lacZ gene in a single replication cycle provided a measure of the retroviral mutation rates. Treatment of D17 target cells with 500 mu M thymidine increased the SNV and MLV mutant frequencies 4.7- and ii-fold, respectively. Treatment of D17 target cells with 2 mM HU increased the SNV and MLV mutant frequencies 2.1- and 2.7-fold, respectively. These results demonstrate that dNTP pool imbalances are associated with an increase in the in vivo retroviral mutation rates, but AZT treatment results in an increase in the retroviral mutation rates by a mechanism not involving alterations in dNTP pools.
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页码:7941 / 7949
页数:9
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共 64 条
  • [1] BEBENEK K, 1989, J BIOL CHEM, V264, P16948
  • [2] BIRION F, 1996, ANTIVIR RES, V29, P111
  • [3] CEPKO CL, 1992, CURRENT PROTOCOLS MO
  • [4] HIV POPULATION-DYNAMICS IN-VIVO - IMPLICATIONS FOR GENETIC-VARIATION, PATHOGENESIS, AND THERAPY
    COFFIN, JM
    [J]. SCIENCE, 1995, 267 (5197) : 483 - 489
  • [5] COHEN A, 1983, J BIOL CHEM, V258, P2334
  • [6] THE SEQUENCE CONTEXT OF THE INITIATION CODON IN THE ENCEPHALOMYOCARDITIS VIRUS LEADER MODULATES EFFICIENCY OF INTERNAL TRANSLATION INITIATION
    DAVIES, MV
    KAUFMAN, RJ
    [J]. JOURNAL OF VIROLOGY, 1992, 66 (04) : 1924 - 1932
  • [7] GENERATION AND CHARACTERIZATION OF A HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 (HIV-1) MUTANT RESISTANT TO AN HIV-1 PROTEASE INHIBITOR
    ELFARRASH, MA
    KURODA, MJ
    KITAZAKI, T
    MASUDA, T
    KATO, K
    HATANAKA, M
    HARADA, S
    [J]. JOURNAL OF VIROLOGY, 1994, 68 (01) : 233 - 239
  • [8] HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 POL GENE-MUTATIONS WHICH CAUSE DECREASED SUSCEPTIBILITY TO 2',3'-DIDEOXYCYTIDINE
    FITZGIBBON, JE
    HOWELL, RM
    HABERZETTL, CA
    SPERBER, SJ
    GOCKE, DJ
    DUBIN, DT
    [J]. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 1992, 36 (01) : 153 - 157
  • [9] EFFECTS OF 3'-AZIDO-3'-DEOXYTHYMIDINE ON THE DEOXYNUCLEOTIDE TRIPHOSPHATE POOLS OF CULTURED HUMAN-CELLS
    FRICK, LW
    NELSON, DJ
    STCLAIR, MH
    FURMAN, PA
    KRENITSKY, TA
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1988, 154 (01) : 124 - 129
  • [10] ENZYMATIC ASSAY FOR QUANTIFICATION OF DEOXYNUCLEOSIDE TRIPHOSPHATES IN HUMAN-CELLS EXPOSED TO ANTIRETROVIRAL 2',3'-DIDEOXYNUCLEOSIDES
    GAO, WY
    JOHNS, DG
    MITSUYA, H
    [J]. ANALYTICAL BIOCHEMISTRY, 1994, 222 (01) : 116 - 122