Simultaneous mutations at Tyr‐181 and Tyr‐188 in HIV‐1 reverse transcriptase prevents inhibition of RNA‐dependent DNA polymerase activity by the bisheteroarylpiperazine (BHAP) U‐90152s

FEBS Letters

Gary Tarpley, W.²  Evans, David B.³  Thomas, Richard C.¹  Fan, Naisheng³  Sharma, Satish K.³  Rank, Kenneth B.³ 

[1] Medicinal Chemistry, Upjohn Laboratories, Kalamazoo, MI 49001, USA;Cancer and Infectious Diseases Research, Upjohn Laboratories, Kalamazoo, MI 49001, USA;Biochemistry, Upjohn Laboratories, Kalamazoo, MI 49001, USA

关键词: HIV-1 reverse transcriptase;    HIV-1 RT mutant;    Resistance to U-90152S;    HIV-2 RT;    Double mutant;    HIV-1;    human immunodeficiency virus type 1;    HIV-2;    human immunodeficiency virus type 2;    RT;    reverse transcriptase;    AIDS;    acquired immunodeficiency syndrome;    IMAC;    immobilized metal affinity chromatography;    SDS;    sodium dodecyl sulfate;    PAGE;    polyacrylamide gel electrophoresis;    BSA;    bovine serum albumin;    TCA;    trichloroacetic acid;    TIBO R82913;    (+)-(5S)-4;    5;    6;    7-tetrahydro-9-chloro-5-methyl-6-(3-methyl-2-butenylimidazo[4;    5;    1-j;    k][1;    4;    ]benzodiazepin-2(1H)- thione;    BHAP;    bisheteroarylpiperazine;    NNRTIs;    nonnucleoside reverse transcriptase inhibitors;    PCR;    polymerase chain reaction;   

DOI  :  10.1016/0014-5793(95)00793-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The replacement of either Tyr-181 or Tyr-188 of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) by the corresponding HIV-2 RT amino acids Ile-181 or Leu-188 is known to result in active mutant enzymes (Y181I; Y188L) with virtual loss of sensitivity towards three structural classes of nonnucleoside RT inhibitors; L-697,661, nevirapine, and TIBO R82913. The bisheteroarylpiperazine (BHAP) U-90152S, a highly specific inhibitor (IC₅₀, 0.29 ± 0.01 μM) of HIV-1 RT, inhibited the recombinant Y181I and Y188L HIV-1 RT mutants with IC₅₀ values of 3.6 ± 0.15 μM and 0.71 ± 0.02 μM, respectively. Construction and in vitro analysis of double mutants Y181I/Y188L and Y181C/Y188L of HIV-1 RT showed > 150-fold resistance to U-90152S. An HIV-2 mutant containing amino acids 176–190 from HIV-1 RT acquired full sensitivity to U-90152S (IC₅₀, 0.29 ± 0.01 μM). It is concluded tha simultaneous mutations at Tyr-181 and Tyr-188 of HIV-1 RT promotes resistance to U-90152S.

【 授权许可】

Unknown