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Proc Natl Acad Sci U S A. 2010 Nov 16;107(46):19736-41. doi: 10.1073/pnas.1014139107. Epub 2010 Nov 1.
2
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Kinetic analysis of PCNA clamp binding and release in the clamp loading reaction catalyzed by Saccharomyces cerevisiae replication factor C.酿酒酵母复制因子C催化的钳加载反应中PCNA钳结合与释放的动力学分析。
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本文引用的文献

1
Analysis of the role of PCNA-DNA contacts during clamp loading.钳夹加载过程中PCNA-DNA相互作用的作用分析。
BMC Struct Biol. 2010 Jan 30;10:3. doi: 10.1186/1472-6807-10-3.
2
Extracting kinetics information from single-molecule fluorescence resonance energy transfer data using hidden markov models.使用隐马尔可夫模型从单分子荧光共振能量转移数据中提取动力学信息。
J Phys Chem B. 2009 Aug 20;113(33):11535-42. doi: 10.1021/jp903831z.
3
Mechanism of ATP-driven PCNA clamp loading by S. cerevisiae RFC.酿酒酵母RFC介导的ATP驱动的PCNA钳加载机制。
J Mol Biol. 2009 May 8;388(3):431-42. doi: 10.1016/j.jmb.2009.03.014. Epub 2009 Mar 13.
4
DNA polymerase delta is highly processive with proliferating cell nuclear antigen and undergoes collision release upon completing DNA.DNA聚合酶δ与增殖细胞核抗原结合时具有高度持续性，并在完成DNA合成后发生碰撞释放。
J Biol Chem. 2008 Oct 24;283(43):29522-31. doi: 10.1074/jbc.M804488200. Epub 2008 Jul 16.
5
Division of labor at the eukaryotic replication fork.真核生物复制叉处的分工。
Mol Cell. 2008 Apr 25;30(2):137-44. doi: 10.1016/j.molcel.2008.02.022.
6
On the mechanism of loading the PCNA sliding clamp by RFC.关于复制因子C（RFC）加载增殖细胞核抗原（PCNA）滑动夹的机制。
Mol Microbiol. 2008 Apr;68(1):216-22. doi: 10.1111/j.1365-2958.2008.06150.x. Epub 2008 Feb 26.
7
Structure of a sliding clamp on DNA.DNA上滑动夹的结构。
Cell. 2008 Jan 11;132(1):43-54. doi: 10.1016/j.cell.2007.11.045.
8
Dynamics of human replication factors in the elongation phase of DNA replication.DNA复制延伸阶段人类复制因子的动力学
Nucleic Acids Res. 2007;35(20):6904-16. doi: 10.1093/nar/gkm822. Epub 2007 Oct 11.
9
The role of fluctuations in tRNA selection by the ribosome.核糖体在tRNA选择过程中波动的作用。
Proc Natl Acad Sci U S A. 2007 Aug 21;104(34):13661-5. doi: 10.1073/pnas.0705988104. Epub 2007 Aug 15.
10
Proliferating cell nuclear antigen loaded onto double-stranded DNA: dynamics, minor groove interactions and functional implications.加载到双链DNA上的增殖细胞核抗原：动力学、小沟相互作用及功能意义
Nucleic Acids Res. 2006;34(20):6023-33. doi: 10.1093/nar/gkl744. Epub 2006 Oct 27.

酵母夹的逐步加载通过整体和单分子研究揭示。

Stepwise loading of yeast clamp revealed by ensemble and single-molecule studies.

机构信息

Department of Chemistry, Pennsylvania State University, University Park, PA 16802, USA.

出版信息

Proc Natl Acad Sci U S A. 2010 Nov 16;107(46):19736-41. doi: 10.1073/pnas.1014139107. Epub 2010 Nov 1.

DOI:10.1073/pnas.1014139107
PMID:21041673
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2993353/
Abstract

In ensemble and single-molecule experiments using the yeast proliferating cell nuclear antigen (PCNA, clamp) and replication factor C (RFC, clamp loader), we have examined the assembly of the RFC·PCNA·DNA complex and its progression to holoenzyme upon addition of polymerase δ (polδ). We obtained data that indicate (i) PCNA loading on DNA proceeds through multiple conformational intermediates and is successful after several failed attempts; (ii) RFC does not act catalytically on a primed 45-mer templated fork; (iii) the RFC·PCNA·DNA complex formed in the presence of ATP is derived from at least two kinetically distinguishable species; (iv) these species disassemble through either unloading of RFC·PCNA from DNA or dissociation of PCNA into its component subunits; and (v) in the presence of polδ only one species converts to the RFC·PCNA·DNA·polδ holoenzyme. These findings redefine and deepen our understanding of the clamp-loading process and reveal that it is surprisingly one of trial and error to arrive at a heuristic solution.

摘要

在使用酵母增殖细胞核抗原 (PCNA，夹子) 和复制因子 C (RFC，夹子加载器) 的组合和单分子实验中，我们研究了 RFC·PCNA·DNA 复合物的组装及其在加入聚合酶 δ (polδ) 后向全酶的进展。我们获得的数据表明：(i) PCNA 在 DNA 上的加载通过多个构象中间体进行，并且在几次失败的尝试后成功；(ii) RFC 对引物 45-mer 模板叉没有催化作用；(iii) 在 ATP 存在下形成的 RFC·PCNA·DNA 复合物来自至少两种动力学上可区分的物种；(iv) 这些物种通过从 DNA 上卸下 RFC·PCNA 或 PCNA 解离成其组成亚基而解体；(v) 在 polδ 的存在下，只有一种物种转化为 RFC·PCNA·DNA·polδ 全酶。这些发现重新定义并加深了我们对夹子加载过程的理解，并揭示了它是一个令人惊讶的反复试验以达到启发式解决方案的过程。