title: PRP19 transforms into a sensor of RPA-ssDNA after DNA damage and drives ATR activation via a ubiquitin-mediated circuitry.
abstract: PRP19 is a ubiquitin ligase involved in pre-mRNA splicing and the DNA damage response (DDR). Although the role for PRP19 in splicing is well characterized, its role in the DDR remains elusive. Through a proteomic screen for proteins that interact with RPA-coated single-stranded DNA (RPA-ssDNA), we identified PRP19 as a sensor of DNA damage. PRP19 directly binds RPA and localizes to DNA damage sites via RPA, promoting RPA ubiquitylation in a DNA-damage-induced manner. PRP19 facilitates the accumulation of ATRIP, the regulatory partner of the ataxia telangiectasia mutated and Rad3-related (ATR) kinase, at DNA damage sites. Depletion of PRP19 compromised the phosphorylation of ATR substrates, recovery of stalled replication forks, and progression of replication forks on damaged DNA. Importantly, PRP19 mutants that cannot bind RPA or function as an E3 ligase failed to support the ATR response, revealing that PRP19 drives ATR activation by acting as an RPA-ssDNA-sensing ubiquitin ligase during the DDR.
authors: Massachusetts General Hospital Cancer Center Harvard Medical School Charlestown, MA 02129.
authors: Alexandre
authors: A
authors: Maréchal
authors: Department of Biochemistry and Molecular Biology The University of Texas Health Science Center at Houston Houston, TX 77030.
authors: Ju-Mei
authors: JM
authors: Li
authors: Massachusetts General Hospital Cancer Center Harvard Medical School Charlestown, MA 02129.
authors: Xiao Ye
authors: XY
authors: Ji
authors: Massachusetts General Hospital Cancer Center Harvard Medical School Charlestown, MA 02129.
authors: Ching-Shyi
authors: CS
authors: Wu
authors: Massachusetts General Hospital Cancer Center Harvard Medical School Charlestown, MA 02129.
authors: Stephanie A
authors: SA
authors: Yazinski
authors: Massachusetts General Hospital Cancer Center Harvard Medical School Charlestown, MA 02129.
authors: Hai Dang
authors: HD
authors: Nguyen
authors: Massachusetts General Hospital Cancer Center Harvard Medical School Charlestown, MA 02129.
authors: Shizhou
authors: S
authors: Liu
authors: Massachusetts General Hospital Cancer Center Harvard Medical School Charlestown, MA 02129.
authors: Amanda E
authors: AE
authors: Jiménez
authors: Department of Biochemistry and Molecular Biology The University of Texas Health Science Center at Houston Houston, TX 77030.
authors: Jianping
authors: J
authors: Jin
authors: Massachusetts General Hospital Cancer Center Harvard Medical School Charlestown, MA 02129.|Department of Pathology Massachusetts General Hospital Harvard Medical School Boston, MA 02114.
authors: Lee
authors: L
authors: Zou
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D064007:Ataxia Telangiectasia Mutated Proteins / Q000378:metabolism / Q000502:physiology; D000071877:Checkpoint Kinase 1; D004249:DNA Damage; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000502:physiology*; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000502:physiology*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D000072260:RNA Splicing Factors; D051716:Replication Protein A / Q000378:metabolism* / Q000502:physiology; D015398:Signal Transduction; D025801:Ubiquitin / Q000378:metabolism / Q000502:physiology*
chemical_list: C521194:ATRIP protein, human; D048868:Adaptor Proteins, Signal Transducing; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D000072260:RNA Splicing Factors; D051716:Replication Protein A; D025801:Ubiquitin; D011494:Protein Kinases; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D000071877:Checkpoint Kinase 1; D045643:DNA Repair Enzymes; C417041:PRPF19 protein, human