title: Metnase promotes restart and repair of stalled and collapsed replication forks.
abstract: Metnase is a human protein with methylase (SET) and nuclease domains that is widely expressed, especially in proliferating tissues. Metnase promotes non-homologous end-joining (NHEJ), and knockdown causes mild hypersensitivity to ionizing radiation. Metnase also promotes plasmid and viral DNA integration, and topoisomerase IIα (TopoIIα)-dependent chromosome decatenation. NHEJ factors have been implicated in the replication stress response, and TopoIIα has been proposed to relax positive supercoils in front of replication forks. Here we show that Metnase promotes cell proliferation, but it does not alter cell cycle distributions, or replication fork progression. However, Metnase knockdown sensitizes cells to replication stress and confers a marked defect in restart of stalled replication forks. Metnase promotes resolution of phosphorylated histone H2AX, a marker of DNA double-strand breaks at collapsed forks, and it co-immunoprecipitates with PCNA and RAD9, a member of the PCNA-like RAD9-HUS1-RAD1 intra-S checkpoint complex. Metnase also promotes TopoIIα-mediated relaxation of positively supercoiled DNA. Metnase is not required for RAD51 focus formation after replication stress, but Metnase knockdown cells show increased RAD51 foci in the presence or absence of replication stress. These results establish Metnase as a key factor that promotes restart of stalled replication forks, and implicate Metnase in the repair of collapsed forks.
authors.affiliation: Department of Molecular Genetics and Microbiology, Division of Hematology-Oncology, Cancer Research and Treatment Center, University of New Mexico School of Medicine, Albuquerque, NM 87131, USA.
authors.forename: Leyma P
authors.initials: LP
authors.lastname: De Haro
authors.forename: Justin
authors.initials: J
authors.lastname: Wray
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Williamson
authors.forename: Stephen T
authors.initials: ST
authors.lastname: Durant
authors.forename: Lori
authors.initials: L
authors.lastname: Corwin
authors.forename: Amanda C
authors.initials: AC
authors.lastname: Gentry
authors.forename: Neil
authors.initials: N
authors.lastname: Osheroff
authors.forename: Suk-Hee
authors.initials: SH
authors.lastname: Lee
authors.forename: Robert
authors.initials: R
authors.lastname: Hromas
authors.forename: Jac A
authors.initials: JA
authors.lastname: Nickoloff
mesh_terms: D000951:Antigens, Neoplasm / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000302:isolation & purification; D049109:Cell Proliferation; D002470:Cell Survival; D004260:DNA Repair; D004261:DNA Replication; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004278:DNA, Superhelical / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D011495:Histone-Lysine N-Methyltransferase / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D006657:Histones / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D018809:Proliferating Cell Nuclear Antigen / Q000302:isolation & purification; D051135:Rad51 Recombinase / Q000032:analysis
chemical_list: D000951:Antigens, Neoplasm; D018797:Cell Cycle Proteins; D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; C468783:H2AX protein, human; D006657:Histones; D018809:Proliferating Cell Nuclear Antigen; C069930:rad9 protein; D011495:Histone-Lysine N-Methyltransferase; C510450:SETMAR protein, human; D051135:Rad51 Recombinase; D004250:DNA Topoisomerases, Type II
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