Structural requirements for enzymatic activities of foamy virus protease‐reverse transcriptase
Structural requirements for enzymatic activities of foamy virus protease‐reverse transcriptase
复制标题
泡沫病毒蛋白酶逆转录酶酶活性的结构要求
DOI：
10.1002/prot.24394
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Hartl MJ
中科院分区：
文献类型：
--
作者：
Schneider A;Peter D;Schmitt J;Richter F;Rösch P;Wöhrl BM;Hartl MJ
逆转录酶（RT）在逆转录病毒的生命周期中起关键作用，并将基因组病毒RNA转化为双链DNA。RT聚合酶结构域被细分为手指、手掌、拇指和连接亚结构域，连接亚结构域将聚合酶连接到C末端RNA酶H结构域。与正逆转录病毒相反，泡沫病毒的成熟RT在其N末端（PR-RT）具有蛋白酶（PR）结构域。因此，由于与其他RT的同源性低，很难定义（子）结构域的边界和功能。我们将N-和C-末端缺失引入猴泡沫病毒PR-RT中，以研究截短对催化活性的影响。RNA酶H结构域和连接亚结构域都对聚合酶的完整性和稳定性以及聚合酶活性和底物结合有很大贡献。PR的42个氨基酸长的C末端区域对于聚合酶的稳定性和活性是重要的。PR激活通过PR-RT与病毒RNA的结合需要全长PR-RT（包括RNase H结构域）的存在。在体外，FV PR对Gag和Pol切割位点的切割效率相当，即使在病毒颗粒中，只有Pol位点被完全切割，这表明其他因素控制PR活性，病毒成熟需要严格调节。Proteins 2014; 82：375-385.© 2013 Wiley Periodicals，Inc.
Reverse transcriptases (RTs) are pivotal in the life cycle of retroviruses and convert the genomic viral RNA into double‐stranded DNA. The RT polymerase domain is subdivided into fingers, palm, thumb, and the connection subdomain, which links the polymerase to the C‐terminal RNase H domain. In contrast to orthoretroviruses, mature RT of foamy viruses harbors the protease (PR) domain at its N‐terminus (PR‐RT). Therefore and due to low homology to other RTs, it is difficult to define the boundaries and functions of the (sub)domains. We introduced N‐ and C‐terminal deletions into simian foamy virus PR‐RT to investigate the impact of the truncations on the catalytic activities. Both, the RNase H domain and the connection subdomain contribute substantially to polymerase integrity and stability as well as to polymerase activity and substrate binding. The 42 amino acids long region C‐terminal of the PR is important for polymerase stability and activity. PR activationviabinding of PR‐RT to viral RNA requires the presence of the full length PR‐RT including the RNase H domain.In vitro, the cleavage efficiencies of FV PR for the Gag and Pol cleavage site are comparable, even though in virus particles only the Pol site is cleaved to completion suggesting that additional factors control PR activity and that virus maturation needs to be strictly regulated. Proteins 2014; 82:375–385. © 2013 Wiley Periodicals, Inc.
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