Characterization of the human immunodeficiency virus type-1 reverse transcriptase enzyme produced in yeast.
Characterization of the human immunodeficiency virus type-1 reverse transcriptase enzyme produced in yeast.
复制标题
酵母中产生的人类免疫缺陷病毒 1 型逆转录酶的表征。
DOI：
10.1016/0006-291x(90)91187-w
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发表时间：
1990
影响因子：
3.1
通讯作者：
Barr,PJ
中科院分区：
文献类型：
--
作者：
Bathurst,IC;Moen,LK;Lujan,MA;Gibson,HL;Feucht,PH;Pichuantes,S;Craik,CS;Santi,DV;Barr,PJ
人类免疫缺陷病毒1型（human immunodeficiency virus type-1，HIV-1）的逆转录酶（reverse transcriptase，RT）由分子量约为66 kD和51 kD的两个亚基组成。我们利用酵母表达的66 kD RT和HIV-1蛋白酶（PR）确定了51 kD分子的羧基端。精确的构建体编码的66 kD和51 kD分子单独表达，在酵母中，在高水平。纯化的重组亚基被证明是关联成异二聚体，保留RT和RNase H活性。只有66 kD分子才能结合成同源二聚体。此类同二聚体保留了异二聚体约80%的RT活性。我们的数据表明，51 - 66 kD的异源二聚体，类似于体内发现的，可以在体外重建，是更有效的RT和RNA酶H活性比同源二聚体。
The reverse transcriptase (RT) of human immunodeficiency virus type-1 (HIV-1) is comprised of two subunits of approximately 66kD and 51kD. We have defined the carboxyl terminus of the 51kD molecule using the 66kD RT and HIV-1 protease (PR) expressed in yeast. Precise constructs encoding the 66kD and 51kD molecules were expressed individually, in yeast, at high levels. The purified recombinant subunits were shown to associate into heterodimers that retained both RT and RNase H activities. Only the 66kD molecule could associate into homodimers. Such homodimers retained approximately 80% of the RT activity of the heterodimers. Our data demonstrates that the 51 66 kD heterodimer, analogous to that found in vivo, can be reconstituted in vitro and is more efficient in both RT and RNase H activity than the homodimer.