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Mos1转座酶-反向重复DNA复合物的结晶:生化及初步晶体学分析

Crystallization of a Mos1 transposase-inverted-repeat DNA complex: biochemical and preliminary crystallographic analyses.

作者信息

Richardson Julia M, Finnegan David J, Walkinshaw Malcolm D

机构信息

School of Biological Sciences, University of Edinburgh, The King's Buildings, Mayfield Road, Edinburgh EH9 3JR, Scotland.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 May 1;63(Pt 5):434-7. doi: 10.1107/S1744309107019045. Epub 2007 Apr 20.

DOI:10.1107/S1744309107019045
PMID:17565190
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2335011/
Abstract

A complex formed between Mos1 transposase and its inverted-repeat DNA has been crystallized. The crystals diffract to 3.25 A resolution and exhibit monoclinic (P2(1)) symmetry, with unit-cell parameters a = 120.8, b = 85.1, c = 131.6 A, beta = 99.3 degrees . The X-ray diffraction data display noncrystallographic twofold symmetry and characteristic dsDNA diffraction at approximately 3.3 A. Biochemical analyses confirmed the presence of DNA and full-length protein in the crystals. The relationship between the axis of noncrystallographic symmetry, the unit-cell axes and the DNA diffraction pattern are discussed. The data are consistent with the previously proposed model of the paired-ends complex containing a dimer of the transposase.

摘要

已使Mos1转座酶与其反向重复DNA形成的复合物结晶。这些晶体的衍射分辨率达到3.25埃,并呈现单斜(P2(1))对称性,晶胞参数为a = 120.8、b = 85.1、c = 131.6埃,β = 99.3°。X射线衍射数据显示非晶体学二重对称性以及在约3.3埃处的特征性双链DNA衍射。生化分析证实晶体中存在DNA和全长蛋白质。讨论了非晶体学对称轴、晶胞轴与DNA衍射图谱之间的关系。这些数据与先前提出的包含转座酶二聚体的配对末端复合物模型一致。

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本文引用的文献

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Mol Ther. 2007 Jan;15(1):139-45. doi: 10.1038/sj.mt.6300028.
2
Transposons for gene therapy!用于基因治疗的转座子!
Curr Gene Ther. 2006 Oct;6(5):593-607. doi: 10.2174/156652306778520647.
3
Mechanism of Mos1 transposition: insights from structural analysis.Mos1转座机制:结构分析带来的见解
EMBO J. 2006 Mar 22;25(6):1324-34. doi: 10.1038/sj.emboj.7601018. Epub 2006 Mar 2.
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Molecular architecture of a eukaryotic DNA transposase.真核生物DNA转座酶的分子结构
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5
Assembly of the mariner Mos1 synaptic complex.水手Mos1突触复合体的组装。
Mol Cell Biol. 2005 Apr;25(7):2861-70. doi: 10.1128/MCB.25.7.2861-2870.2005.
6
Mos as a tool for genome-wide insertional mutagenesis in Caenorhabditis elegans: results of a pilot study.Mos作为秀丽隐杆线虫全基因组插入诱变的工具:一项初步研究的结果。
Nucleic Acids Res. 2004 Aug 13;32(14):e117. doi: 10.1093/nar/gnh111.
7
Structural analysis of the bipartite DNA-binding domain of Tc3 transposase bound to transposon DNA.与转座子DNA结合的Tc3转座酶二分体DNA结合结构域的结构分析。
Nucleic Acids Res. 2004 Aug 10;32(14):4306-12. doi: 10.1093/nar/gkh770. Print 2004.
8
Expression, purification and preliminary crystallographic studies of a single-point mutant of Mos1 mariner transposase.Mos1 水手转座酶单点突变体的表达、纯化及初步晶体学研究
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