A mechanism of AZT resistance: An increase in nucleotide-dependent primer unblocking by mutant HIV-1 reverse transcriptase

被引:287
|
作者
Meyer, PR
Matsuura, SE
Mian, AM
So, AG
Scott, WA [1 ]
机构
[1] Univ Miami, Sch Med, Dept Biochem & Mol Biol, Miami, FL 33101 USA
[2] Univ Miami, Sch Med, Dept Med, Miami, FL 33101 USA
关键词
D O I
10.1016/S1097-2765(00)80185-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Mutations in HIV-1 reverse transcriptase (RT) give rise to 3'-azido-3'-deoxythymidine (AZT) resistance by a mechanism that has not been previously reproduced in vitro. We show that mutant RT has increased ability to remove AZTMP from blocked primers through a nucleotide-dependent reaction, producing dinucleoside polyphosphate and extendible primer. In the presence of physiological concentrations of ATP, mutant RT extended 12% to 15% of primers past multiple AZTMP termination sites versus less than 0.5% for wild type. Although mutant RT also unblocked ddAMP-terminated primers more efficiently than wild-type RT, the removal of ddAMP was effectively inhibited by the next complementary dNTP (ICS50 approximate to 12 mu M). In contrast, the removal of AZTMP was not inhibited by dNTPs except at nonphysiological concentrations (ICS50 > 200 mu M).
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页码:35 / 43
页数:9
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