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RecO 重组介体蛋白的晶体结构来自 和其与 DNA 和锌离子的相互作用。

Crystal Structure of the Recombination Mediator Protein RecO from and Its Interaction with DNA and a Zinc Ion.

机构信息

Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea.

Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon 24341, Korea.

出版信息

Int J Mol Sci. 2022 Aug 26;23(17):9667. doi: 10.3390/ijms23179667.

DOI:10.3390/ijms23179667
PMID:36077065
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9456098/
Abstract

Homologous recombination is involved in repairing DNA damage, contributing to maintaining the integrity and stability of viral and cellular genomes. In bacteria, the recombination mediator proteins RecO and RecR are required to load the RecA recombinase on ssDNA for homologous recombination. To structurally and functionally characterize RecO, we determined the crystal structure of RecO from (cjRecO) at a 1.8 Å resolution and biochemically assessed its capacity to interact with DNA and a metal ion. cjRecO folds into a curved rod-like structure that consists of an N-terminal domain (NTD), C-terminal domain (CTD), and Zn-binding domain (ZnD). The ZnD at the end of the rod-like structure coordinates three cysteine residues and one histidine residue to accommodate a Zn ion. Based on an extensive comparative analysis of RecO structures and sequences, we propose that the Zn-binding consensus sequence of RecO is CxxC…C/HxxC/H/D. The interaction with Zn is indispensable for the protein stability of cjRecO but does not seem to be required for the recombination mediator function. cjRecO also interacts with ssDNA as part of its biological function, potentially using the positively charged patch in the NTD and CTD. However, cjRecO displays a low ssDNA-binding affinity, suggesting that cjRecO requires RecR to efficiently recognize ssDNA for homologous recombination.

摘要

同源重组参与修复 DNA 损伤，有助于维持病毒和细胞基因组的完整性和稳定性。在细菌中，重组介体蛋白 RecO 和 RecR 被需要将 RecA 重组酶加载到 ssDNA 上以进行同源重组。为了对 RecO 进行结构和功能表征，我们确定了来自 (cjRecO) 的晶体结构，分辨率为 1.8Å，并从生物化学上评估了其与 DNA 和金属离子相互作用的能力。cjRecO 折叠成一个弯曲的杆状结构，由 N 端结构域 (NTD)、C 端结构域 (CTD) 和 Zn 结合结构域 (ZnD) 组成。杆状结构末端的 ZnD 协调三个半胱氨酸残基和一个组氨酸残基以容纳 Zn 离子。基于对 RecO 结构和序列的广泛比较分析，我们提出 RecO 的 Zn 结合保守序列为 CxxC…C/HxxC/H/D。与 Zn 的相互作用对于 cjRecO 的蛋白质稳定性是必不可少的，但似乎不需要重组介体功能。cjRecO 还作为其生物学功能的一部分与 ssDNA 相互作用，可能使用 NTD 和 CTD 中的正电荷斑。然而，cjRecO 显示出低的 ssDNA 结合亲和力，表明 cjRecO 需要 RecR 来有效地识别 ssDNA 以进行同源重组。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/6395015b2d6d/ijms-23-09667-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/8c134e0b245a/ijms-23-09667-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/455cf5576673/ijms-23-09667-g002.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/eae44c1470a8/ijms-23-09667-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/6395015b2d6d/ijms-23-09667-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/8c134e0b245a/ijms-23-09667-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/455cf5576673/ijms-23-09667-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/b68f83fd1dc7/ijms-23-09667-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/eae44c1470a8/ijms-23-09667-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/577a/9456098/6395015b2d6d/ijms-23-09667-g005.jpg

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Mismatch sensing by nucleofilament deciphers mechanism of RecA-mediated homologous recombination.
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