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古菌 FANCM 同源物 Hef 对停滞的 DNA 复制的细胞内动力学。

Intracellular dynamics of archaeal FANCM homologue Hef in response to halted DNA replication.

机构信息

Laboratoire d'Optique et Biosciences, Ecole Polytechnique, CNRS UMR7645-INSERM U696, 91128 Palaiseau Cedex, France and Section of Molecular Cytology, Swammerdam Institute for Life Sciences, van Leeuwenhoek Centre for Advanced Microscopy, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands.

出版信息

Nucleic Acids Res. 2013 Dec;41(22):10358-70. doi: 10.1093/nar/gkt816. Epub 2013 Sep 17.

DOI:10.1093/nar/gkt816
PMID:24049073
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3905845/
Abstract

Hef is an archaeal member of the DNA repair endonuclease XPF (XPF)/Crossover junction endonuclease MUS81 (MUS81)/Fanconi anemia, complementation group M (FANCM) protein family that in eukaryotes participates in the restart of stalled DNA replication forks. To investigate the physiological roles of Hef in maintaining genome stability in living archaeal cells, we studied the localization of Hef-green fluorescent protein fusions by fluorescence microscopy. Our studies revealed that Haloferax volcanii Hef proteins formed specific localization foci under regular growth conditions, the number of which specifically increased in response to replication arrest. Purification of the full-length Hef protein from its native host revealed that it forms a stable homodimer in solution, with a peculiar elongated configuration. Altogether our data indicate that the shape of Hef, significant physicochemical constraints and/or interactions with DNA limit the apparent cytosolic diffusion of halophilic DNA replication/repair complexes, and demonstrate that Hef proteins are dynamically recruited to archaeal eukaryotic-like chromatin to counteract DNA replication stress. We suggest that the evolutionary conserved function of Hef/FANCM proteins is to enhance replication fork stability by directly interacting with collapsed replication forks.

摘要

Hef 是一种古菌成员，属于 DNA 修复内切酶 XPF（XPF）/ 交叉连接内切酶 MUS81（MUS81）/ 范可尼贫血症互补组 M（FANCM）蛋白家族，在真核生物中参与停滞 DNA 复制叉的重新启动。为了研究 Hef 在维持活古菌细胞基因组稳定性中的生理作用，我们通过荧光显微镜研究了 Hef-绿色荧光蛋白融合物的定位。我们的研究表明，在常规生长条件下，Haloferax volcanii Hef 蛋白形成特定的定位焦点，其数量在复制停滞时特异性增加。从其天然宿主中纯化全长 Hef 蛋白表明，它在溶液中形成稳定的同源二聚体，具有奇特的伸长构象。总之，我们的数据表明，Hef 的形状、显著的物理化学限制和/或与 DNA 的相互作用限制了嗜盐 DNA 复制/修复复合物的明显细胞质扩散，并表明 Hef 蛋白被动态募集到古菌真核样染色质以抵消 DNA 复制应激。我们认为，Hef/FANCM 蛋白的进化保守功能是通过直接与崩溃的复制叉相互作用来增强复制叉的稳定性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/e2d7f4aa30ca/gkt816f7p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/8686a654dba0/gkt816f1p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/3514aa74973b/gkt816f2p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/b3920933388f/gkt816f3p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/10354d759bdb/gkt816f4p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/faebddfedc2b/gkt816f5p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/3fabaa5a43ea/gkt816f6p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/e2d7f4aa30ca/gkt816f7p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/8686a654dba0/gkt816f1p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/3514aa74973b/gkt816f2p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/b3920933388f/gkt816f3p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/10354d759bdb/gkt816f4p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/faebddfedc2b/gkt816f5p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/3fabaa5a43ea/gkt816f6p.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01a5/3905845/e2d7f4aa30ca/gkt816f7p.jpg

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