TY - JOUR
T1 - PARG is essential for Polθ-mediated DNA end-joining by removing repressive poly-ADP-ribose marks
AU - Vekariya, Umeshkumar
AU - Minakhin, Leonid
AU - Chandramouly, Gurushankar
AU - Tyagi, Mrityunjay
AU - Kent, Tatiana
AU - Sullivan-Reed, Katherine
AU - Atkins, Jessica
AU - Ralph, Douglas
AU - Nieborowska-Skorska, Margaret
AU - Kukuyan, Anna Mariya
AU - Tang, Hsin Yao
AU - Pomerantz, Richard T.
AU - Skorski, Tomasz
N1 - Publisher Copyright:
© The Author(s) 2024.
PY - 2024/7/11
Y1 - 2024/7/11
N2 - DNA polymerase theta (Polθ)-mediated end-joining (TMEJ) repairs DNA double-strand breaks and confers resistance to genotoxic agents. How Polθ is regulated at the molecular level to exert TMEJ remains poorly characterized. We find that Polθ interacts with and is PARylated by PARP1 in a HPF1-independent manner. PARP1 recruits Polθ to the vicinity of DNA damage via PARylation dependent liquid demixing, however, PARylated Polθ cannot perform TMEJ due to its inability to bind DNA. PARG-mediated de-PARylation of Polθ reactivates its DNA binding and end-joining activities. Consistent with this, PARG is essential for TMEJ and the temporal recruitment of PARG to DNA damage corresponds with TMEJ activation and dissipation of PARP1 and PAR. In conclusion, we show a two-step spatiotemporal mechanism of TMEJ regulation. First, PARP1 PARylates Polθ and facilitates its recruitment to DNA damage sites in an inactivated state. PARG subsequently activates TMEJ by removing repressive PAR marks on Polθ.
AB - DNA polymerase theta (Polθ)-mediated end-joining (TMEJ) repairs DNA double-strand breaks and confers resistance to genotoxic agents. How Polθ is regulated at the molecular level to exert TMEJ remains poorly characterized. We find that Polθ interacts with and is PARylated by PARP1 in a HPF1-independent manner. PARP1 recruits Polθ to the vicinity of DNA damage via PARylation dependent liquid demixing, however, PARylated Polθ cannot perform TMEJ due to its inability to bind DNA. PARG-mediated de-PARylation of Polθ reactivates its DNA binding and end-joining activities. Consistent with this, PARG is essential for TMEJ and the temporal recruitment of PARG to DNA damage corresponds with TMEJ activation and dissipation of PARP1 and PAR. In conclusion, we show a two-step spatiotemporal mechanism of TMEJ regulation. First, PARP1 PARylates Polθ and facilitates its recruitment to DNA damage sites in an inactivated state. PARG subsequently activates TMEJ by removing repressive PAR marks on Polθ.
KW - Poly ADP Ribosylation
KW - Poly Adenosine Diphosphate Ribose/metabolism
KW - DNA/metabolism
KW - DNA Polymerase theta
KW - DNA-Directed DNA Polymerase/metabolism
KW - Poly(ADP-ribose) Polymerases/metabolism
KW - Humans
KW - DNA Breaks, Double-Stranded
KW - DNA-Binding Proteins/metabolism
KW - Nuclear Proteins
KW - Animals
KW - Glycoside Hydrolases
KW - DNA End-Joining Repair
KW - HEK293 Cells
KW - Poly (ADP-Ribose) Polymerase-1/metabolism
KW - DNA Damage
KW - Carrier Proteins
UR - http://www.scopus.com/inward/record.url?scp=85198134063&partnerID=8YFLogxK
U2 - 10.1038/s41467-024-50158-7
DO - 10.1038/s41467-024-50158-7
M3 - Article
C2 - 38987289
AN - SCOPUS:85198134063
SN - 2041-1723
VL - 15
SP - 5822
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 5822
ER -