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

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

Author information

1
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.

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.

PMID:
24049073
PMCID:
PMC3905845
DOI:
10.1093/nar/gkt816
[Indexed for MEDLINE]
Free PMC Article

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