Common determinants in DNA melting and helicase-catalysed DNA unwinding by papillomavirus replication protein E1.

Common determinants in DNA melting and helicase-catalysed DNA unwinding by papillomavirus replication protein E1.
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DOI:
10.1093/nar/gkl384
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发表时间:
2006
影响因子:
14.9
通讯作者:
Sanders CM
Sanders CM
中科院分区:
生物学2区
文献类型:
--
作者:
Castella S;Bingham G;Sanders CM

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肿瘤病毒牛乳头瘤病毒(BPV-1)和猿猴病毒40 (SV40)的E1和t抗原是在DNA复制初始阶段识别和融化各自复制起点的启动蛋白。然后这些蛋白质在单链DNA上组装成六聚体解旋酶。在t抗原中,一个特有的环和发夹结构(前传感器1β发夹,PS1βH)投射到由蛋白质六聚体化产生的中心空腔中。这个通道经历了很大的atp依赖的构象变化,并且环路/PS1βH被认为形成了一个对解旋酶活性至关重要的DNA结合位点。在这里,我们发现BPV E1中的保守残基可能形成类似的环状/发夹结构,这是解旋酶活性和起源(或)DNA熔化所必需的。我们提出DNA的融化需要E1解旋酶结构域(E1HD)和DNA上的起源结合结构域(OBD)的合作。一种可能的机制是,当DNA锁定在环/PS1βH DNA结合位点时,atp依赖的构象变化将DNA以扭转运动向内吸引,以促进解绕。
E1 and T-antigen of the tumour viruses bovine papillomavirus (BPV-1) and Simian virus 40 (SV40) are the initiator proteins that recognize and melt their respective origins of replication in the initial phase of DNA replication. These proteins then assemble into processive hexameric helicases upon the single-stranded DNA that they create. In T-antigen, a characteristic loop and hairpin structure (the pre-sensor 1β hairpin, PS1βH) project into a central cavity generated by protein hexamerization. This channel undergoes large ATP-dependent conformational changes, and the loop/PS1βH is proposed to form a DNA binding site critical for helicase activity. Here, we show that conserved residues in BPV E1 that probably form a similar loop/hairpin structure are required for helicase activity and also origin (ori) DNA melting. We propose that DNA melting requires the cooperation of the E1 helicase domain (E1HD) and the origin binding domain (OBD) tethered to DNA. One possible mechanism is that with the DNA locked in the loop/PS1βH DNA binding site, ATP-dependent conformational changes draw the DNA inwards in a twisting motion to promote unwinding.
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