DNA damage-induced ubiquitylation of RFC2 subunit of replication factor C complex

Junya Tomida, Yuji Masuda, Hidekazu Hiroaki, Tomoko Ishikawa, Ihnyoung Song, Toshiki Tsurimoto, Satoshi Tateishi, Tadahiro Shiomi, Yasuhiro Kamei, Jinhyeong Kim, Kenji Kamiya, Cyrus Vaziri, Haruo Ohmori, Takeshi Todo

研究成果: ジャーナルへの寄稿記事

20 引用 (Scopus)

抄録

Many proteins involved in DNA replication and repair undergo post-translational modifications such as phosphorylation and ubiquitylation. Proliferating cell nuclear antigen (PCNA; a homotrimeric protein that encircles double-stranded DNA to function as a sliding clamp for DNA polymerases) is monoubiquitylated by the RAD6-RAD18 complex and further polyubiquitylated by the RAD5-MMS2-UBC13 complex in response to various DNA-damaging agents. PCNA mono- and polyubiquitylation activate an error-prone translesion synthesis pathway and an error-free pathway of damage avoidance, respectively. Here we show that replication factor C (RFC; a heteropentameric protein complex that loads PCNA onto DNA) was also ubiquitylated in a RAD18-dependent manner in cells treated with alkylating agents or H2O2. A mutant form of RFC2 with a D228A substitution (corresponding to a yeast Rfc4 mutation that reduces an interaction with replication protein A (RPA), a single-stranded DNA-binding protein) was heavily ubiquitylated in cells even in the absence of DNA damage. Furthermore RFC2 was ubiquitylated by the RAD6-RAD18 complex in vitro, and its modification was inhibited in the presence of RPA. The inhibitory effect of RPA on RFC2 ubiquitylation was relatively specific because RAD6-RAD18-mediated ubiquitylation of PCNA was RPA-insensitive. Our findings suggest that RPA plays a regulatory role in DNA damage responses via repression of RFC2 ubiquitylation in human cells.

元の言語英語
ページ(範囲)9071-9079
ページ数9
ジャーナルJournal of Biological Chemistry
283
発行部数14
DOI
出版物ステータス出版済み - 4 4 2008

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Replication Protein C
Replication Protein A
Ubiquitination
Proliferating Cell Nuclear Antigen
DNA Damage
DNA
Proteins
Alkylating Agents
DNA-Binding Proteins
DNA-Directed DNA Polymerase
Post Translational Protein Processing
Phosphorylation
DNA Replication
DNA Repair
Clamping devices
Yeast
Yeasts
Repair
Substitution reactions
Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

DNA damage-induced ubiquitylation of RFC2 subunit of replication factor C complex. / Tomida, Junya; Masuda, Yuji; Hiroaki, Hidekazu; Ishikawa, Tomoko; Song, Ihnyoung; Tsurimoto, Toshiki; Tateishi, Satoshi; Shiomi, Tadahiro; Kamei, Yasuhiro; Kim, Jinhyeong; Kamiya, Kenji; Vaziri, Cyrus; Ohmori, Haruo; Todo, Takeshi.

:: Journal of Biological Chemistry, 巻 283, 番号 14, 04.04.2008, p. 9071-9079.

研究成果: ジャーナルへの寄稿記事

Tomida, J, Masuda, Y, Hiroaki, H, Ishikawa, T, Song, I, Tsurimoto, T, Tateishi, S, Shiomi, T, Kamei, Y, Kim, J, Kamiya, K, Vaziri, C, Ohmori, H & Todo, T 2008, 'DNA damage-induced ubiquitylation of RFC2 subunit of replication factor C complex', Journal of Biological Chemistry, 巻. 283, 番号 14, pp. 9071-9079. https://doi.org/10.1074/jbc.M709835200
Tomida, Junya ; Masuda, Yuji ; Hiroaki, Hidekazu ; Ishikawa, Tomoko ; Song, Ihnyoung ; Tsurimoto, Toshiki ; Tateishi, Satoshi ; Shiomi, Tadahiro ; Kamei, Yasuhiro ; Kim, Jinhyeong ; Kamiya, Kenji ; Vaziri, Cyrus ; Ohmori, Haruo ; Todo, Takeshi. / DNA damage-induced ubiquitylation of RFC2 subunit of replication factor C complex. :: Journal of Biological Chemistry. 2008 ; 巻 283, 番号 14. pp. 9071-9079.
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AU - Tomida, Junya

AU - Masuda, Yuji

AU - Hiroaki, Hidekazu

AU - Ishikawa, Tomoko

AU - Song, Ihnyoung

AU - Tsurimoto, Toshiki

AU - Tateishi, Satoshi

AU - Shiomi, Tadahiro

AU - Kamei, Yasuhiro

AU - Kim, Jinhyeong

AU - Kamiya, Kenji

AU - Vaziri, Cyrus

AU - Ohmori, Haruo

AU - Todo, Takeshi

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N2 - Many proteins involved in DNA replication and repair undergo post-translational modifications such as phosphorylation and ubiquitylation. Proliferating cell nuclear antigen (PCNA; a homotrimeric protein that encircles double-stranded DNA to function as a sliding clamp for DNA polymerases) is monoubiquitylated by the RAD6-RAD18 complex and further polyubiquitylated by the RAD5-MMS2-UBC13 complex in response to various DNA-damaging agents. PCNA mono- and polyubiquitylation activate an error-prone translesion synthesis pathway and an error-free pathway of damage avoidance, respectively. Here we show that replication factor C (RFC; a heteropentameric protein complex that loads PCNA onto DNA) was also ubiquitylated in a RAD18-dependent manner in cells treated with alkylating agents or H2O2. A mutant form of RFC2 with a D228A substitution (corresponding to a yeast Rfc4 mutation that reduces an interaction with replication protein A (RPA), a single-stranded DNA-binding protein) was heavily ubiquitylated in cells even in the absence of DNA damage. Furthermore RFC2 was ubiquitylated by the RAD6-RAD18 complex in vitro, and its modification was inhibited in the presence of RPA. The inhibitory effect of RPA on RFC2 ubiquitylation was relatively specific because RAD6-RAD18-mediated ubiquitylation of PCNA was RPA-insensitive. Our findings suggest that RPA plays a regulatory role in DNA damage responses via repression of RFC2 ubiquitylation in human cells.

AB - Many proteins involved in DNA replication and repair undergo post-translational modifications such as phosphorylation and ubiquitylation. Proliferating cell nuclear antigen (PCNA; a homotrimeric protein that encircles double-stranded DNA to function as a sliding clamp for DNA polymerases) is monoubiquitylated by the RAD6-RAD18 complex and further polyubiquitylated by the RAD5-MMS2-UBC13 complex in response to various DNA-damaging agents. PCNA mono- and polyubiquitylation activate an error-prone translesion synthesis pathway and an error-free pathway of damage avoidance, respectively. Here we show that replication factor C (RFC; a heteropentameric protein complex that loads PCNA onto DNA) was also ubiquitylated in a RAD18-dependent manner in cells treated with alkylating agents or H2O2. A mutant form of RFC2 with a D228A substitution (corresponding to a yeast Rfc4 mutation that reduces an interaction with replication protein A (RPA), a single-stranded DNA-binding protein) was heavily ubiquitylated in cells even in the absence of DNA damage. Furthermore RFC2 was ubiquitylated by the RAD6-RAD18 complex in vitro, and its modification was inhibited in the presence of RPA. The inhibitory effect of RPA on RFC2 ubiquitylation was relatively specific because RAD6-RAD18-mediated ubiquitylation of PCNA was RPA-insensitive. Our findings suggest that RPA plays a regulatory role in DNA damage responses via repression of RFC2 ubiquitylation in human cells.

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