title: SUMO-modification and elimination of the active DNA demethylation enzyme TDG in cultured human cells.
abstract: Thymine DNA glycosylase (TDG) is a base excision repair enzyme that interacts with the small ubiquitin-related modifier (SUMO)-targeted ubiquitin E3 ligase RNF4 and functions in the active DNA demethylation pathway. Here we showed that both SUMOylated and non-modified forms of endogenous TDG fluctuated during the cell cycle and in response to drugs that perturbed cell cycle progression, including hydroxyurea and nocodazole. Additionally, we detected a SUMOylation-independent association between TDG and RNF4 in vitro as well as in vivo, and observed that both forms of TDG were efficiently degraded in RNF4-depleted cells when arrested at S phase. Our findings provide insights into the in vivo dynamics of TDG SUMOylation and further clarify the TDG-RNF4 interaction.
authors.affiliation: Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan.
authors.forename: Taishi
authors.initials: T
authors.lastname: Moriyama
authors.affiliation: Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Fujimitsu
authors.affiliation: Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan.
authors.forename: Yushi
authors.initials: Y
authors.lastname: Yoshikai
authors.affiliation: Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Sasano
authors.affiliation: Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Yamada
authors.affiliation: Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan.
authors.forename: Masataka
authors.initials: M
authors.lastname: Murakami
authors.affiliation: Department of Biochemistry, Shimane University School of Medicine, 89-1 Enya-cho, Izumo 693-8501, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Urano
authors.affiliation: Biosignal Research Center, Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada-ku, Kobe 657-8501, Japan.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Sugasawa
authors.affiliation: Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan; Department of New Frontier Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan. Electronic address: hisa@kumamoto-u.ac.jp.
authors.forename: Hisato
authors.initials: H
authors.lastname: Saitoh
mesh_terms: D002453:Cell Cycle / Q000187:drug effects; D002478:Cells, Cultured; D019175:DNA Methylation; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D009154:Mutation; D015739:Nocodazole / Q000494:pharmacology; D009687:Nuclear Proteins / Q000378:metabolism*; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*; D058207:Sumoylation; D044502:Thymine DNA Glycosylase / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D009687:Nuclear Proteins; C114070:RNF4 protein, human; D025841:Small Ubiquitin-Related Modifier Proteins; D014157:Transcription Factors; D044502:Thymine DNA Glycosylase; D015739:Nocodazole; D006918:Hydroxyurea
keywords: Cell cycle; DNA demethylation; Ring finger protein 4 (RNF4); SUMO-targeted ubiquitin ligase (STUbL); Small ubiquitin-related modifier (SUMO); Thymine DNA glycosylase (TDG)