Metnase promotes restart and repair of stalled and collapsed replication forks

Leyma P. de Haro, Justin Wray, Elizabeth A. Williamson, Stephen T. Durant, Lori Corwin, Amanda C. Gentry, Neil Osheroff, Suk Hee Lee, Robert Hromas, Jac A. Nickoloff

Research output: Contribution to journalArticle

42 Scopus citations

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.

Original languageEnglish (US)
Article numbergkq339
Pages (from-to)5681-5691
Number of pages11
JournalNucleic acids research
Volume38
Issue number17
DOIs
StatePublished - May 10 2010
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

Fingerprint Dive into the research topics of 'Metnase promotes restart and repair of stalled and collapsed replication forks'. Together they form a unique fingerprint.

  • Cite this

    de Haro, L. P., Wray, J., Williamson, E. A., Durant, S. T., Corwin, L., Gentry, A. C., Osheroff, N., Lee, S. H., Hromas, R., & Nickoloff, J. A. (2010). Metnase promotes restart and repair of stalled and collapsed replication forks. Nucleic acids research, 38(17), 5681-5691. [gkq339]. https://doi.org/10.1093/nar/gkq339