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一种宿主编码的DNA结合蛋白在特定序列的复制终点促进质粒复制的终止。

A host-encoded DNA-binding protein promotes termination of plasmid replication at a sequence-specific replication terminus.

作者信息

Sista P R, Mukherjee S, Patel P, Khatri G S, Bastia D

机构信息

Department of Microbiology and Immunology, Duke University Medical Center, Durham, NC 27710.

出版信息

Proc Natl Acad Sci U S A. 1989 May;86(9):3026-30. doi: 10.1073/pnas.86.9.3026.

Abstract

We have purified approximately 6600-fold an approximately 40-kDa protein (Ter protein) encoded by Escherichia coli that specifically binds to two sites at the 216-base-pair replication terminus (tau) of the plasmid R6K. Chemical footprinting experiments have shown that the Ter protein binds to two 14- to 16-base-pair sequences that exist as inverted repeats in the tau fragment. Site-directed mutagenesis of one of the terminus sequences (tau R) resulted in a mutant tau R that failed to bind to the Ter protein. The same mutant terminus also failed to terminate DNA replication in vivo. These experiments strongly suggest that the interaction of the Ter protein with tau sequences plays an essential role in the termination of DNA replication, specifically at tau.

摘要

我们已经将大肠杆菌编码的一种约40 kDa的蛋白质(Ter蛋白)纯化了约6600倍,该蛋白质特异性结合于质粒R6K的216碱基对复制 terminus(tau)的两个位点。化学足迹实验表明,Ter蛋白结合于tau片段中以反向重复形式存在的两个14至16碱基对序列。对其中一个末端序列(tau R)进行定点诱变,产生了一个无法与Ter蛋白结合的突变体tau R。相同的突变末端在体内也无法终止DNA复制。这些实验有力地表明,Ter蛋白与tau序列的相互作用在DNA复制的终止中起着至关重要的作用,特别是在tau处。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03b9/287057/de15eb3421ed/pnas00249-0046-a.jpg

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