The FHA and BRCT domains recognize ADP-ribosylation during DNA damage response

Abstract

Poly-ADP-ribosylation is a unique post-translational modification participating in many biological processes, such as DNA damage response. Here, we demonstrate that a set of Forkhead-associated (FHA) and BRCA1 C-terminal (BRCT) domains recognizes poly(ADP-ribose) (PAR) both in vitro and in vivo. Among these FHA and BRCT domains, the FHA domains of APTX and PNKP interact with iso-ADP-ribose, the linkage of PAR, whereas the BRCT domains of Ligase4, XRCC1, and NBS1 recognize ADP-ribose, the basic unit of PAR. The interactions between PAR and the FHA or BRCT domains mediate the relocation of these domain-containing proteins to DNA damage sites and facilitate the DNA damage response. Moreover, the interaction between PAR and the NBS1 BRCT domain is important for the early activation of ATM during DNA damage response and ATM-dependent cell cycle checkpoint activation. Taken together, our results demonstrate two novel PAR-binding modules that play important roles in DNA damage response.

The oligonucleotide/oligosaccharide-binding fold motif is a poly(ADP-ribose)-binding domain that mediates DNA damage response

“…S9A). This is consistent with other findings that PAR also mediates the fast recruitment of other DNA damage repair factors to DNA lesions (15,16). To examine the nature of the relatively fast DNA damage repair mediated by hSSB1, we used siRNA to deplete XRCC1, a key factor for ssDNA damage repair (33-37).…”

“…Thus, INTS3 associates with other partners at DNA lesions to stabilize the hSSB1-INTS3 complex. This biological setting is very similar to the BRCA1/BARD1 complex and the FHA/BRCT domain of NBS1, in which one module interacts with PAR for the fast recruitment to DNA damage sites, whereas the other module is responsible for the prolonged retention at DNA lesions (15,16). Both activities are important for DNA damage repair.…”

“…Recent evidence suggests that DNA damage-induced PAR may serve as a docking signal to recruit DDR factors to DNA lesions (7)(8)(9)(10)(11)(12)(13)(14). For example, our recent study showed that the BRCT domains of BARD1 and NBS1 recognize PAR, which facilitates the rapid recruitment of the BRCA1/BARD1 complex and NBS1 to the site of DNA damage (15,16). These results indicate that other DDR factors may also be recruited to DNA damage sites by PAR.…”

Differential and Concordant Roles for Poly(ADP-Ribose) Polymerase 1 and Poly(ADP-Ribose) in Regulating WRN and RECQL5 Activities

“…A third module has been described as a PAR binding zinc finger motif (51). Furthermore, for proteins with WWE, BRCT, or FHA domains, it has been demonstrated that their recruitment to sites of DNA damage depends on PAR binding (13,54). Examination of the primary sequence of each of the human RecQ helicases reveals that they have multiple versions of the PBM-type motif, and the motifs of RECQL5 are shown in Fig.…”

“…This docking site serves to integrate diverse cellular signaling pathways, including DNA damage detection and repair, transcription, chromatin remodeling, and cell death (11). PAR formation plays a critical role in the rapid recruitment of DNA repair proteins such as MRE11 (12), NBS1 (13), BRCA1 (14), and ATM (15) to DSBs. Mice lacking PARP1 are extremely sensitive to genotoxic stresses, show defects in the repair of damaged DNA, and display genomic instability, with high frequencies of sister chromatid exchange and chromosomal breakage (16).…”