Draper Bonnie, Rao Venigalla B
Department of Biology, The Catholic University of America, Washington, DC 20064, USA.
J Mol Biol. 2007 May 25;369(1):79-94. doi: 10.1016/j.jmb.2007.03.019. Epub 2007 Mar 15.
Tailed bacteriophages and large eukaryotic viruses employ powerful molecular motors to translocate dsDNA into a preassembled capsid shell. The phage T4 motor is composed of a dodecameric portal and small and large terminase subunits assembled at the special head-tail connector vertex of the prohead. The motor pumps DNA through the portal channel, utilizing ATP hydrolysis energy provided by an ATPase present in the large terminase subunit. We report that the ATPase motors of terminases, helicases, translocating restriction enzymes, and protein translocases possess a common coupling motif (C-motif). Mutations in the phage T4 terminase C-motif lead to loss of stimulated ATPase and DNA translocation activities. Surprisingly, the mutants can catalyze at least one ATP hydrolysis event but are unable to turn over and reset the motor. This is the first report of a catalytic block in translocating ATPase motor after ATP hydrolysis occurred. We suggest that the C-motif is an ATP hydrolysis sensor, linking product release to mechanical motion. A novel terminase-driven mechanism is proposed for translocation of dsDNA in viruses.
有尾噬菌体和大型真核病毒利用强大的分子马达将双链DNA转运到预先组装好的衣壳壳内。噬菌体T4马达由一个十二聚体门户以及在原头部特殊的头尾连接顶点组装的小和大的末端酶亚基组成。该马达利用大末端酶亚基中存在的ATP酶提供的ATP水解能量,通过门户通道泵送DNA。我们报道,末端酶、解旋酶、转运限制酶和蛋白质转运酶的ATP酶马达具有一个共同的偶联基序(C基序)。噬菌体T4末端酶C基序中的突变导致刺激的ATP酶和DNA转运活性丧失。令人惊讶的是,这些突变体可以催化至少一次ATP水解事件,但无法翻转和重置马达。这是在ATP水解发生后转运ATP酶马达中催化阻断的首次报道。我们认为C基序是一个ATP水解传感器,将产物释放与机械运动联系起来。提出了一种由末端酶驱动的新型机制用于病毒中双链DNA的转运。