title: Thymine DNA glycosylase is a CRL4Cdt2 substrate.
abstract: The E3 ubiquitin ligase CRL4(Cdt2) targets proteins for destruction in S phase and after DNA damage by coupling ubiquitylation to DNA-bound proliferating cell nuclear antigen (PCNA). Coupling to PCNA involves a PCNA-interacting peptide (PIP) degron motif in the substrate that recruits CRL4(Cdt2) while binding to PCNA. In vertebrates, CRL4(Cdt2) promotes degradation of proteins whose presence in S phase is deleterious, including Cdt1, Set8, and p21. Here, we show that CRL4(Cdt2) targets thymine DNA glycosylase (TDG), a base excision repair enzyme that is involved in DNA demethylation. TDG contains a conserved and nearly perfect match to the PIP degron consensus. TDG is ubiquitylated and destroyed in a PCNA-, Cdt2-, and PIP degron-dependent manner during DNA repair in Xenopus egg extract. The protein can also be destroyed during DNA replication in this system. During Xenopus development, TDG first accumulates during gastrulation, and its expression is down-regulated by CRL4(Cdt2). Our results expand the group of vertebrate CRL4(Cdt2) substrates to include a bona fide DNA repair enzyme.
authors.affiliation: Departments of Biological Chemistry and Molecular Pharmacology and Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Tamara J
authors.initials: TJ
authors.lastname: Slenn
authors.affiliation: Departments of Biological Chemistry and Molecular Pharmacology and Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Morris
authors.affiliation: Departments of Biological Chemistry and Molecular Pharmacology and Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Courtney G
authors.initials: CG
authors.lastname: Havens
authors.affiliation: Departments of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Freeman
authors.affiliation: Graduate School of Science, Osaka University, Osaka, Japan, and.
authors.forename: Tatsuro S
authors.initials: TS
authors.lastname: Takahashi
authors.affiliation: Departments of Biological Chemistry and Molecular Pharmacology and Harvard Medical School, Boston, Massachusetts 02115; Howard Hughes Medical Institute, Harvard University, Boston, Massachusetts 02115. Electronic address: johannes_walter@hms.harvard.edu.
authors.forename: Johannes C
authors.initials: JC
authors.lastname: Walter
mesh_terms: D000818:Animals; D019175:DNA Methylation / Q000502:physiology*; D005775:Gastrula / Q000166:cytology / Q000201:enzymology*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D044502:Thymine DNA Glycosylase / Q000378:metabolism*; D043743:Ubiquitin-Protein Ligase Complexes; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination / Q000502:physiology*; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis
chemical_list: D029867:Xenopus Proteins; C552111:dtl protein, Xenopus; D043743:Ubiquitin-Protein Ligase Complexes; D044767:Ubiquitin-Protein Ligases; D044502:Thymine DNA Glycosylase
keywords: Base Excision Repair (BER); CRL4-Cdt2; DNA Methylation; DNA Replication; E3 Ubiquitin Ligase; Proteolysis; Thymine DNA Glycosylase (TDG); Xenopus