HIV Drug Resistance Program, Center for Cancer Research, National Cancer Institute, National Institutes of Health, PO Box B, Frederick, MD 21702, USA.
Virology. 2013 Sep;444(1-2):241-9. doi: 10.1016/j.virol.2013.06.017. Epub 2013 Jul 11.
Previous work on mutations in the thumb of HIV-1 reverse transcriptase (RT) showed that the majority of the mutant RTs were degraded (by the viral protease) to various extents in virions. This degradation was, in most cases, temperature sensitive, and presumably was due to a partial unfolding of the protein at 37°C. We used recombinant proteins to investigate the effects of the mutations on the thermal stability and proteolytic degradation of RT. Both subunits contribute to the stability of RT. In general, the differences in stability between the mutants and WT were greater if the mutation was in p51 rather than p66. Expressing the Pol polyprotein containing the RT mutants in Escherichia coli produced results similar to what was seen in virions; the mutant RTs were misfolded and/or degraded at 37°C, but were better folded and processed at 30°C.
先前有关 HIV-1 逆转录酶（RT）拇指突变的研究表明，大多数突变 RT 在病毒粒子中不同程度地被（病毒蛋白酶）降解。这种降解在大多数情况下是温度敏感的，推测是由于蛋白质在 37°C 时部分展开。我们使用重组蛋白来研究突变对 RT 热稳定性和蛋白水解降解的影响。两个亚基都有助于 RT 的稳定性。一般来说，如果突变发生在 p51 而不是 p66，突变和 WT 之间的稳定性差异更大。在大肠杆菌中表达含有 RT 突变的 Pol 多蛋白产生的结果与病毒粒子中观察到的结果相似；突变 RT 在 37°C 时发生错误折叠和/或降解，但在 30°C 时更好地折叠和加工。
