丙型肝炎病毒NS3解旋酶解开DNA的弹簧加载机制。
Spring-loaded mechanism of DNA unwinding by hepatitis C virus NS3 helicase.
作者信息
Myong Sua, Bruno Michael M, Pyle Anna M, Ha Taekjip
机构信息
Physics Department, University of Illinois, 1110 West Green Street, Urbana, IL 61801, USA.
出版信息
Science. 2007 Jul 27;317(5837):513-6. doi: 10.1126/science.1144130.
Abstract
NS3, an essential helicase for replication of hepatitis C virus, is a model enzyme for investigating helicase function. Using single-molecule fluorescence analysis, we showed that NS3 unwinds DNA in discrete steps of about three base pairs (bp). Dwell time analysis indicated that about three hidden steps are required before a 3-bp step is taken. Taking into account the available structural data, we propose a spring-loaded mechanism in which several steps of one nucleotide per adenosine triphosphate molecule accumulate tension on the protein-DNA complex, which is relieved periodically via a burst of 3-bp unwinding. NS3 appears to shelter the displaced strand during unwinding, and, upon encountering a barrier or after unwinding >18 bp, it snaps or slips backward rapidly and repeats unwinding many times in succession. Such repetitive unwinding behavior over a short stretch of duplex may help to keep secondary structures resolved during viral genome replication.
摘要
NS3是丙型肝炎病毒复制所必需的解旋酶，是研究解旋酶功能的模型酶。通过单分子荧光分析，我们发现NS3以约三个碱基对（bp）的离散步骤解开DNA。驻留时间分析表明，在进行3-bp步骤之前大约需要三个隐藏步骤。考虑到现有的结构数据，我们提出了一种弹簧加载机制，其中每个三磷酸腺苷分子的一个核苷酸的几个步骤在蛋白质-DNA复合物上积累张力，该张力通过3-bp解旋的爆发周期性地释放。NS3在解旋过程中似乎会保护被置换的链，并且在遇到障碍或解旋超过18 bp后，它会迅速向后折断或滑动，并连续多次重复解旋。在短双链体上的这种重复解旋行为可能有助于在病毒基因组复制过程中保持二级结构的解析。
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