Cdt1-binding protein GRWD1 is a novel histone-binding protein that facilitates MCM loading through its influence on chromatin architecture. Issue 12 (18th May 2015)

• Record Type: Journal Article
• Authors: Sugimoto, Nozomi; Maehara, Kazumitsu; Yoshida, Kazumasa; Yasukouchi, Shuhei; Osano, Satoko; Watanabe, Shinya; Aizawa, Masahiro; Yugawa, Takashi; Kiyono, Tohru; Kurumizaka, Hitoshi; Ohkawa, Yasuyuki; Fujita, Masatoshi
• Is Part Of: Nucleic acids research. Volume 43:Issue 12(2015)
• Page Start: 5898
• Page End: 5911
• Publication Date: 2015-05-18
• Subjects: Nucleic acids -- Periodicals; Molecular biology -- Periodicals; 572.805
• DOI: 10.1093/nar/gkv509
• Languages: English
• ISSNs: 0305-1048

Abstract

Abstract: Efficient pre-replication complex (pre-RC) formation on chromatin templates is crucial for the maintenance of genome integrity. However, the regulation of chromatin dynamics during this process has remained elusive. We found that a conserved protein, GRWD1 (glutamate-rich WD40 repeat containing 1), binds to two representative replication origins specifically during G1 phase in a CDC6- and Cdt1-dependent manner, and that depletion of GRWD1 reduces loading of MCM but not CDC6 and Cdt1. Furthermore, chromatin immunoprecipitation coupled with high-throughput sequencing (Seq) revealed significant genome-wide co-localization of GRWD1 with CDC6. We found that GRWD1 has histone-binding activity. To investigate the effect of GRWD1 on chromatin architecture, we used formaldehyde-assisted isolation of regulatory elements (FAIRE)-seq or FAIRE-quantitative PCR analyses, and the results suggest that GRWD1 regulates chromatin openness at specific chromatin locations. Taken together, these findings suggest that GRWD1 may be a novel histone-binding protein that regulates chromatin dynamics and MCM loading at replication origins.