• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

通过人类线粒体DNA聚合酶辅助亚基的结构、建模和诱变揭示的一种新型DNA合成持续合成机制。

A novel processive mechanism for DNA synthesis revealed by structure, modeling and mutagenesis of the accessory subunit of human mitochondrial DNA polymerase.

作者信息

Fan Li, Kim Sangbumn, Farr Carol L, Schaefer Kevin T, Randolph Kathleen M, Tainer John A, Kaguni Laurie S

机构信息

Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA; Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92034, USA.

出版信息

J Mol Biol. 2006 May 19;358(5):1229-43. doi: 10.1016/j.jmb.2006.02.073. Epub 2006 Mar 15.

DOI:10.1016/j.jmb.2006.02.073
PMID:16574152
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4703138/
Abstract

Mitochondrial DNA polymerase (pol gamma) is the sole DNA polymerase responsible for replication and repair of animal mitochondrial DNA. Here, we address the molecular mechanism by which the human holoenzyme achieves high processivity in nucleotide polymerization. We have determined the crystal structure of human pol gamma-beta, the accessory subunit that binds with high affinity to the catalytic core, pol gamma-alpha, to stimulate its activity and enhance holoenzyme processivity. We find that human pol gamma-beta shares a high level of structural similarity to class IIa aminoacyl tRNA synthetases, and forms a dimer in the crystal. A human pol gamma/DNA complex model was developed using the structures of the pol gamma-beta dimer and the bacteriophage T7 DNA polymerase ternary complex, which suggests multiple regions of subunit interaction between pol gamma-beta and the human catalytic core that allow it to encircle the newly synthesized double-stranded DNA, and thereby enhance DNA binding affinity and holoenzyme processivity. Biochemical properties of a novel set of human pol gamma-beta mutants are explained by and test the model, and elucidate the role of the accessory subunit as a novel type of processivity factor in stimulating pol gamma activity and in enhancing processivity.

摘要

线粒体DNA聚合酶（polγ）是负责动物线粒体DNA复制和修复的唯一DNA聚合酶。在此，我们探讨了人类全酶在核苷酸聚合过程中实现高持续合成能力的分子机制。我们确定了人类polγ-β的晶体结构，它是与催化核心polγ-α高亲和力结合的辅助亚基，以刺激其活性并增强全酶的持续合成能力。我们发现人类polγ-β与IIa类氨酰tRNA合成酶具有高度的结构相似性，并在晶体中形成二聚体。利用polγ-β二聚体和噬菌体T7 DNA聚合酶三元复合物的结构建立了人类polγ/DNA复合物模型，该模型表明polγ-β与人类催化核心之间存在多个亚基相互作用区域，使其能够环绕新合成的双链DNA，从而增强DNA结合亲和力和全酶的持续合成能力。一组新型人类polγ-β突变体的生化特性通过该模型得到解释并对其进行了验证，阐明了辅助亚基作为一种新型持续合成因子在刺激polγ活性和增强持续合成能力方面的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/de82369c2ade/nihms747243f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/d0b2aafb352b/nihms747243f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/8d4d8b972777/nihms747243f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/885c2887a2ca/nihms747243f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/86a477cc57ea/nihms747243f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/e8fe0c2d75fb/nihms747243f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/77a43424fdde/nihms747243f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/de82369c2ade/nihms747243f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/d0b2aafb352b/nihms747243f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/8d4d8b972777/nihms747243f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/885c2887a2ca/nihms747243f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/86a477cc57ea/nihms747243f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/e8fe0c2d75fb/nihms747243f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/77a43424fdde/nihms747243f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/325c/4703138/de82369c2ade/nihms747243f7.jpg

相似文献

1
A novel processive mechanism for DNA synthesis revealed by structure, modeling and mutagenesis of the accessory subunit of human mitochondrial DNA polymerase.通过人类线粒体DNA聚合酶辅助亚基的结构、建模和诱变揭示的一种新型DNA合成持续合成机制。
J Mol Biol. 2006 May 19;358(5):1229-43. doi: 10.1016/j.jmb.2006.02.073. Epub 2006 Mar 15.
2
The accessory subunit of mtDNA polymerase shares structural homology with aminoacyl-tRNA synthetases: implications for a dual role as a primer recognition factor and processivity clamp.线粒体DNA聚合酶的辅助亚基与氨酰-tRNA合成酶具有结构同源性：对其作为引物识别因子和持续合成钳的双重作用的启示。
Proc Natl Acad Sci U S A. 1999 Aug 17;96(17):9527-32. doi: 10.1073/pnas.96.17.9527.
3
Synergistic Effects of the T251I and P587L Mitochondrial DNA Polymerase γ Disease Mutations.线粒体DNA聚合酶γ疾病突变T251I和P587L的协同效应
J Biol Chem. 2017 Mar 10;292(10):4198-4209. doi: 10.1074/jbc.M116.773341. Epub 2017 Feb 2.
4
Crystal structure and deletion analysis show that the accessory subunit of mammalian DNA polymerase gamma, Pol gamma B, functions as a homodimer.晶体结构和缺失分析表明，哺乳动物DNA聚合酶γ的辅助亚基PolγB以同二聚体形式发挥作用。
Mol Cell. 2001 Jan;7(1):43-54. doi: 10.1016/s1097-2765(01)00153-8.
5
Structural basis for processivity and antiviral drug toxicity in human mitochondrial DNA replicase.人类线粒体DNA复制酶持续性和抗病毒药物毒性的结构基础
EMBO J. 2015 Jul 14;34(14):1959-70. doi: 10.15252/embj.201591520. Epub 2015 Jun 8.
6
Structural insight into processive human mitochondrial DNA synthesis and disease-related polymerase mutations.对连续性人类线粒体DNA合成及疾病相关聚合酶突变的结构洞察
Cell. 2009 Oct 16;139(2):312-24. doi: 10.1016/j.cell.2009.07.050.
7
DNA polymerase gamma, the mitochondrial replicase.DNA聚合酶γ，即线粒体复制酶。
Annu Rev Biochem. 2004;73:293-320. doi: 10.1146/annurev.biochem.72.121801.161455.
8
A single mutation in human mitochondrial DNA polymerase Pol gammaA affects both polymerization and proofreading activities of only the holoenzyme.人类线粒体 DNA 聚合酶 Pol γA 中的单个突变仅影响全酶的聚合和校对活性。
J Biol Chem. 2010 Sep 3;285(36):28105-16. doi: 10.1074/jbc.M110.122283. Epub 2010 May 31.
9
Multiple regions of subunit interaction in Drosophila mitochondrial DNA polymerase: three functional domains in the accessory subunit.果蝇线粒体DNA聚合酶中多个亚基相互作用区域：辅助亚基中的三个功能结构域。
Biochemistry. 2001 Apr 17;40(15):4780-91. doi: 10.1021/bi010102h.
10
Each monomer of the dimeric accessory protein for human mitochondrial DNA polymerase has a distinct role in conferring processivity.二聚体辅助蛋白的每个单体在赋予其延伸性方面都有独特的作用。
J Biol Chem. 2010 Jan 8;285(2):1490-9. doi: 10.1074/jbc.M109.062752. Epub 2009 Oct 26.

引用本文的文献

1
Activating AMPK improves pathological phenotypes due to mtDNA depletion.激活AMPK可改善因线粒体DNA耗竭所致的病理表型。
FEBS J. 2025 May;292(9):2359-2380. doi: 10.1111/febs.70006. Epub 2025 Feb 7.
2
Mechanisms and pathologies of human mitochondrial DNA replication and deletion formation.人类线粒体 DNA 复制和缺失形成的机制和病理学。
Biochem J. 2024 Jun 5;481(11):683-715. doi: 10.1042/BCJ20230262.
3
Structural basis for DNA proofreading.DNA 校对的结构基础。

本文引用的文献

1
Processing of X-ray diffraction data collected in oscillation mode.振荡模式下收集的X射线衍射数据的处理。
Methods Enzymol. 1997;276:307-26. doi: 10.1016/S0076-6879(97)76066-X.
2
Functional human mitochondrial DNA polymerase gamma forms a heterotrimer.功能性人类线粒体DNA聚合酶γ形成一个异源三聚体。
J Biol Chem. 2006 Jan 6;281(1):374-82. doi: 10.1074/jbc.M509730200. Epub 2005 Nov 1.
3
The common A467T mutation in the human mitochondrial DNA polymerase (POLG) compromises catalytic efficiency and interaction with the accessory subunit.
Nat Commun. 2023 Dec 27;14(1):8501. doi: 10.1038/s41467-023-44198-8.
4
Structure-specific roles for PolG2-DNA complexes in maintenance and replication of mitochondrial DNA.PolG2-DNA 复合物在维持和复制线粒体 DNA 中的结构特异性作用。
Nucleic Acids Res. 2023 Oct 13;51(18):9716-9732. doi: 10.1093/nar/gkad679.
5
Polγ coordinates DNA synthesis and proofreading to ensure mitochondrial genome integrity.聚合酶 γ 协调 DNA 合成和校对以确保线粒体基因组的完整性。
Nat Struct Mol Biol. 2023 Jun;30(6):812-823. doi: 10.1038/s41594-023-00980-2. Epub 2023 May 18.
6
Structural and Molecular Basis for Mitochondrial DNA Replication and Transcription in Health and Antiviral Drug Toxicity.线粒体 DNA 复制和转录的结构和分子基础：健康与抗病毒药物毒性
Molecules. 2023 Feb 14;28(4):1796. doi: 10.3390/molecules28041796.
7
Automated systematic evaluation of cryo-EM specimens with SmartScope.利用 SmartScope 对冷冻电镜标本进行自动化系统评估。
Elife. 2022 Aug 23;11:e80047. doi: 10.7554/eLife.80047.
8
Method for the structural analysis of Twinkle mitochondrial DNA helicase by cryo-EM.利用冷冻电镜技术对 Twinkle 线粒体 DNA 解旋酶进行结构分析的方法。
Methods. 2022 Sep;205:263-270. doi: 10.1016/j.ymeth.2022.06.012. Epub 2022 Jun 30.
9
TWINKLE and Other Human Mitochondrial DNA Helicases: Structure, Function and Disease.TWINKLE与其他人类线粒体DNA解旋酶：结构、功能与疾病
Genes (Basel). 2020 Apr 9;11(4):408. doi: 10.3390/genes11040408.
10
Replicative DNA polymerases promote active displacement of SSB proteins during lagging strand synthesis.复制 DNA 聚合酶在滞后链合成过程中促进 SSB 蛋白的主动置换。
Nucleic Acids Res. 2019 Jun 20;47(11):5723-5734. doi: 10.1093/nar/gkz249.
人类线粒体DNA聚合酶（POLG）中常见的A467T突变会损害催化效率以及与辅助亚基的相互作用。
J Biol Chem. 2005 Sep 9;280(36):31341-6. doi: 10.1074/jbc.M506762200. Epub 2005 Jul 16.
4
Computation of electrostatic forces between solvated molecules determined by the Poisson-Boltzmann equation using a boundary element method.使用边界元法通过泊松-玻尔兹曼方程计算溶剂化分子之间的静电力。
J Chem Phys. 2005 Jun 1;122(21):214102. doi: 10.1063/1.1924448.
5
Cellular DNA replicases: components and dynamics at the replication fork.细胞DNA复制酶：复制叉处的组分与动态变化
Annu Rev Biochem. 2005;74:283-315. doi: 10.1146/annurev.biochem.73.011303.073859.
6
Functional defects due to spacer-region mutations of human mitochondrial DNA polymerase in a family with an ataxia-myopathy syndrome.一个患有共济失调-肌病综合征的家族中，人类线粒体DNA聚合酶间隔区突变导致的功能缺陷。
Hum Mol Genet. 2005 Jul 15;14(14):1907-20. doi: 10.1093/hmg/ddi196. Epub 2005 May 25.
7
Mutations in the spacer region of Drosophila mitochondrial DNA polymerase affect DNA binding, processivity, and the balance between Pol and Exo function.果蝇线粒体DNA聚合酶间隔区的突变会影响DNA结合、持续合成能力以及聚合酶和外切酶功能之间的平衡。
J Biol Chem. 2005 Jan 28;280(4):2491-7. doi: 10.1074/jbc.M411447200. Epub 2004 Nov 10.
8
Structure and function of the double-strand break repair machinery.双链断裂修复机制的结构与功能
DNA Repair (Amst). 2004 Aug-Sep;3(8-9):863-73. doi: 10.1016/j.dnarep.2004.03.022.
9
Structure-function defects of human mitochondrial DNA polymerase in autosomal dominant progressive external ophthalmoplegia.常染色体显性进行性眼外肌麻痹中人类线粒体DNA聚合酶的结构-功能缺陷
Nat Struct Mol Biol. 2004 Aug;11(8):770-6. doi: 10.1038/nsmb805. Epub 2004 Jul 18.
10
DNA polymerase gamma, the mitochondrial replicase.DNA聚合酶γ，即线粒体复制酶。
Annu Rev Biochem. 2004;73:293-320. doi: 10.1146/annurev.biochem.72.121801.161455.