PARG is essential for Polθ-mediated DNA end-joining by removing repressive poly-ADP-ribose marks

Umeshkumar Vekariya; Leonid Minakhin; Gurushankar Chandramouly; Mrityunjay Tyagi; Tatiana Kent; Katherine Sullivan-Reed; Jessica Atkins; Douglas Ralph; Margaret Nieborowska-Skorska; Anna Mariya Kukuyan; Hsin Yao Tang; Richard T. Pomerantz; Tomasz Skorski

doi:10.1038/s41467-024-50158-7

PARG is essential for Polθ-mediated DNA end-joining by removing repressive poly-ADP-ribose marks

Umeshkumar Vekariya
, Leonid Minakhin
, Gurushankar Chandramouly
, Mrityunjay Tyagi
, Tatiana Kent
, Katherine Sullivan-Reed
, Jessica Atkins
, Douglas Ralph
, Margaret Nieborowska-Skorska
, Anna Mariya Kukuyan
, Hsin Yao Tang
, Richard T. Pomerantz
, Tomasz Skorski

Research output: Contribution to journal › Article › peer-review

18 Scopus citations

Abstract

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.

Original language	English
Article number	5822
Pages (from-to)	5822
Journal	Nature Communications
Volume	15
Issue number	1
DOIs	https://doi.org/10.1038/s41467-024-50158-7
State	Published - Jul 11 2024

Keywords

Poly ADP Ribosylation
Poly Adenosine Diphosphate Ribose/metabolism
DNA/metabolism
DNA Polymerase theta
DNA-Directed DNA Polymerase/metabolism
Poly(ADP-ribose) Polymerases/metabolism
Humans
DNA Breaks, Double-Stranded
DNA-Binding Proteins/metabolism
Nuclear Proteins
Animals
Glycoside Hydrolases
DNA End-Joining Repair
HEK293 Cells
Poly (ADP-Ribose) Polymerase-1/metabolism
DNA Damage
Carrier Proteins

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10.1038/s41467-024-50158-7

Cite this

Vekariya, U., Minakhin, L., Chandramouly, G., Tyagi, M., Kent, T., Sullivan-Reed, K., Atkins, J., Ralph, D., Nieborowska-Skorska, M., Kukuyan, A. M., Tang, H. Y., Pomerantz, R. T., & Skorski, T. (2024). PARG is essential for Polθ-mediated DNA end-joining by removing repressive poly-ADP-ribose marks. Nature Communications, 15(1), 5822. Article 5822. https://doi.org/10.1038/s41467-024-50158-7

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title = "PARG is essential for Polθ-mediated DNA end-joining by removing repressive poly-ADP-ribose marks",

abstract = "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.",

keywords = "Poly ADP Ribosylation, Poly Adenosine Diphosphate Ribose/metabolism, DNA/metabolism, DNA Polymerase theta, DNA-Directed DNA Polymerase/metabolism, Poly(ADP-ribose) Polymerases/metabolism, Humans, DNA Breaks, Double-Stranded, DNA-Binding Proteins/metabolism, Nuclear Proteins, Animals, Glycoside Hydrolases, DNA End-Joining Repair, HEK293 Cells, Poly (ADP-Ribose) Polymerase-1/metabolism, DNA Damage, Carrier Proteins",

author = "Umeshkumar Vekariya and Leonid Minakhin and Gurushankar Chandramouly and Mrityunjay Tyagi and Tatiana Kent and Katherine Sullivan-Reed and Jessica Atkins and Douglas Ralph and Margaret Nieborowska-Skorska and Kukuyan, \{Anna Mariya\} and Tang, \{Hsin Yao\} and Pomerantz, \{Richard T.\} and Tomasz Skorski",

note = "Publisher Copyright: {\textcopyright} The Author(s) 2024.",

year = "2024",

month = jul,

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doi = "10.1038/s41467-024-50158-7",

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Vekariya, U, Minakhin, L, Chandramouly, G, Tyagi, M, Kent, T, Sullivan-Reed, K, Atkins, J, Ralph, D, Nieborowska-Skorska, M, Kukuyan, AM, Tang, HY, Pomerantz, RT & Skorski, T 2024, 'PARG is essential for Polθ-mediated DNA end-joining by removing repressive poly-ADP-ribose marks', Nature Communications, vol. 15, no. 1, 5822, pp. 5822. https://doi.org/10.1038/s41467-024-50158-7

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 - https://www.scopus.com/pages/publications/85198134063

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 -

PARG is essential for Polθ-mediated DNA end-joining by removing repressive poly-ADP-ribose marks

Abstract

Keywords

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Research on Science Detailed by Researchers at Temple University (PARG is essential for Polth-mediated DNA end-joining by removing repressive poly-ADP-ribose marks)

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PARG is essential for Polθ-mediated DNA end-joining by removing repressive poly-ADP-ribose marks

Abstract

Keywords

Access to Document

Other files and links

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Research on Science Detailed by Researchers at Temple University (PARG is essential for Polth-mediated DNA end-joining by removing repressive poly-ADP-ribose marks)

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