TIMELESS Forms a Complex with PARP1 Distinct from Its Complex with TIPIN and Plays a Role in the DNA Damage Response

• Published in: Cell reports (Cambridge) Vol. 13; no. 3; pp. 451 - 459
• Main Authors: Young, Lauren M., Marzio, Antonio, Perez-Duran, Pablo, Reid, Dylan A., Meredith, Daniel N., Roberti, Domenico, Star, Ayelet, Rothenberg, Eli, Ueberheide, Beatrix, Pagano, Michele
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.10.2015; Elsevier
• Subjects: Amino Acid Sequence; Binding Sites; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell Cycle Proteins - chemistry; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cell Line, Tumor; DNA Damage; DNA Repair; DNA-Binding Proteins; HEK293 Cells; Humans; Intracellular Signaling Peptides and Proteins - chemistry; Intracellular Signaling Peptides and Proteins - genetics; Intracellular Signaling Peptides and Proteins - metabolism; Molecular Sequence Data; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerases - genetics; Poly(ADP-ribose) Polymerases - metabolism; Protein Binding
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2015.09.017

PARP1 is the main sensor of single- and double-strand breaks in DNA and, in building chains of poly(ADP-ribose), promotes the recruitment of many downstream signaling and effector proteins involved in the DNA damage response (DDR). We show a robust physical interaction between PARP1 and the replication fork protein TIMELESS, distinct from the known TIMELESS-TIPIN complex, which activates the intra-S phase checkpoint. TIMELESS recruitment to laser-induced sites of DNA damage is dependent on its binding to PARP1, but not PARP1 activity. We also find that the PARP1-TIMELESS complex contains a number of established PARP1 substrates, and TIMELESS mutants unable to bind PARP1 are impaired in their ability to bind PARP1 substrates. Further, PARP1 binding to certain substrates and their recruitment to DNA damage lesions is impaired by TIMELESS knockdown, and TIMELESS silencing significantly impairs DNA double-strand break repair. We hypothesize that TIMELESS cooperates in the PARP1-mediated DDR. [Display omitted] •TIMELESS forms a complex with PARP1 distinct from the TIMELESS-TIPIN complex•TIMELESS is recruited to laser-induced DNA damage sites•TIMELESS recruitment is dependent on PARP1 binding, but not PARP1 activity•TIMELESS binds PARP1 substrates involved in the DNA damage response TIMELESS interacts with TIPIN to control the intra-S phase checkpoint. Young et al. reveal a robust interaction of TIMELESS with PARP1, which is distinct from its interaction with TIPIN. TIMELESS is recruited to laser-induced DNA damage sites in a PARP1-dependent but PARP1-activity-independent manner and participates in the DNA damage response.