title: PHRF1 promotes genome integrity by modulating non-homologous end-joining.
abstract: Methylated histone readers are critical for chromatin dynamics, transcription, and DNA repair. Human PHRF1 contains a plant homeodomain (PHD) that recognizes methylated histones and a RING domain, which ubiquitinates substrates. A recent study reveals that PHRF1 is a tumor suppressor that promotes TGF-β cytostatic signaling through TGIF ubiquitination. Also, PHRF1 is a putative phosphorylation substrate of ataxia telangiectasia-mutated/ataxia telangiectasia and Rad3-related kinases; however, the role of PHRF1 in DNA damage response is unclear. Here we report a novel function of PHRF1 in modulating non-homologous end-joining (NHEJ). PHRF1 quickly localizes to DNA damage lesions upon genotoxic insults. Ablation of PHRF1 decreases the efficiency of plasmid-based end-joining, whereas PHRF1 overexpression leads to an elevated NHEJ in H1299 reporter cells. Immunoprecipitation and peptide pull-down assays verify that PHRF1 constitutively binds to di- and trimethylated histone H3 lysine 36 (H3K36) (H3K36me2 and H3K36me3) via its PHD domain. Substitution of S915DT917E to ADAE in PHRF1 decreases its affinity for NBS1. Both PHD domain and SDTE motif are required for its NHEJ-promoting activity. Furthermore, PHRF1 mediates PARP1 polyubiquitination for proteasomal degradation. These results suggest that PHRF1 may combine with H3K36 methylation and NBS1 to promote NHEJ and stabilize genomic integrity upon DNA damage insults.
authors: Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.
authors: C-F
authors: CF
authors: Chang
authors: Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.
authors: P-C
authors: PC
authors: Chu
authors: Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.
authors: P-Y
authors: PY
authors: Wu
authors: Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 11529, Taiwan.
authors: M-Y
authors: MY
authors: Yu
authors: Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 11529, Taiwan.
authors: J-Y
authors: JY
authors: Lee
authors: 1] Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan [2] Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 11529, Taiwan.
authors: M-D
authors: MD
authors: Tsai
authors: 1] Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan [2] Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 11529, Taiwan.
authors: M-S
authors: MS
authors: Chang
mesh_terms: D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D004249:DNA Damage; D059766:DNA End-Joining Repair; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D015894:Genome, Human; D057809:HEK293 Cells; D006657:Histones / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008745:Methylation; D008969:Molecular Sequence Data; D063146:Polycomb-Group Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D006657:Histones; D018398:Homeodomain Proteins; C112309:PHF1 protein, human; D063146:Polycomb-Group Proteins