title: Usp7-dependent histone H3 deubiquitylation regulates maintenance of DNA methylation.
abstract: Uhrf1-dependent histone H3 ubiquitylation plays a crucial role in the maintenance of DNA methylation via the recruitment of the DNA methyltransferase Dnmt1 to DNA methylation sites. However, the involvement of deubiquitylating enzymes (DUBs) targeting ubiquitylated histone H3 in the maintenance of DNA methylation is largely unknown. With the use of Xenopus egg extracts, we demonstrate here that Usp7, a ubiquitin carboxyl-terminal hydrolase, forms a stable complex with Dnmt1 and is recruited to DNA methylation sites during DNA replication. Usp7 deubiquitylates ubiquitylated histone H3 in vitro. Inhibition of Usp7 activity or its depletion in egg extracts results in enhanced and extended binding of Dnmt1 to chromatin, suppressing DNA methylation. Depletion of Usp7 in HeLa cells causes enhanced histone H3 ubiquitylation and enlargement of Dnmt1 nuclear foci during DNA replication. Our results thus suggest that Usp7 is a key factor that regulates maintenance of DNA methylation.
authors.affiliation: Department of Cell Biology, Graduate School of Medical Sciences, Nagoya City University, 1, Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, 467-8601, Japan.
authors.forename: Luna
authors.initials: L
authors.lastname: Yamaguchi
authors.affiliation: Division of Cancer Cell biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo, 108-8639, Japan. anishiya@ims.u-tokyo.ac.jp.
authors.forename: Atsuya
authors.initials: A
authors.lastname: Nishiyama
authors.affiliation: Department of Cell Biology, Graduate School of Medical Sciences, Nagoya City University, 1, Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, 467-8601, Japan.
authors.forename: Toshinori
authors.initials: T
authors.lastname: Misaki
authors.affiliation: Division of Cancer Cell biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo, 108-8639, Japan.
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Johmura
authors.affiliation: Center for Education in Laboratory Animal Research, Chubu University, Kasugai, Aichi, 487-8501, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Ueda
authors.affiliation: Graduate School of Medical Life Sciences, Yokohama City University, 1-7-29, Suehiro-cho, Tsurumi-ku, Yokohama, 230-0045, Japan.
authors.forename: Kyohei
authors.initials: K
authors.lastname: Arita
authors.affiliation: Division of Molecular Life Science, Graduate School of Life Science, Hokkaido University, Sapporo, 001-0020, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Nagao
authors.affiliation: Division of Molecular Life Science, Graduate School of Life Science, Hokkaido University, Sapporo, 001-0020, Japan.
authors.forename: Chikashi
authors.initials: C
authors.lastname: Obuse
authors.affiliation: Department of Cell Biology, Graduate School of Medical Sciences, Nagoya City University, 1, Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, 467-8601, Japan. mkt-naka@ims.u-tokyo.ac.jp.|Division of Cancer Cell biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo, 108-8639, Japan. mkt-naka@ims.u-tokyo.ac.jp.
authors.forename: Makoto
authors.identifier: 0000-0002-6707-3584
authors.initials: M
authors.lastname: Nakanishi
mesh_terms: D000818:Animals; D002843:Chromatin / Q000378:metabolism; D000076123:DNA (Cytosine-5-)-Methyltransferase 1 / Q000378:metabolism; D019175:DNA Methylation; D004261:DNA Replication / Q000502:physiology; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D010063:Ovum; D000076084:Ubiquitin-Specific Peptidase 7 / Q000378:metabolism*; D054875:Ubiquitination; D014981:Xenopus
chemical_list: D002843:Chromatin; D006657:Histones; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D000076084:Ubiquitin-Specific Peptidase 7