Site-specific acetylation of polynucleotide kinase 3'-phosphatase regulates its distinct role in DNA repair pathways

Islam, Azharul; Chakraborty, Anirban; Sarker, Altaf H.; Aryal, Uma K.; Pan, Lang; Sharma, Gulshan; Boldogh, Istvan; Hazra, Tapas

doi:10.1093/nar/gkae002

Site-specific acetylation of polynucleotide kinase 3'-phosphatase regulates its distinct role in DNA repair pathways

Journal Article · Mon Jan 15 04:00:00 EST 2024 · Nucleic Acids Research

DOI:https://doi.org/10.1093/nar/gkae002· OSTI ID:2470684

Islam, Azharul ^[1]; Chakraborty, Anirban ^[1]; Sarker, Altaf H. ^[2]; Aryal, Uma K. ^[3]; Pan, Lang ^[1]; Sharma, Gulshan ^[1]; Boldogh, Istvan ^[1]; ^[1]

University of Texas Medical Branch, Galveston, TX (United States)
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Purdue Univ., West Lafayette, IN (United States)

Mammalian polynucleotide kinase 3'-phosphatase (PNKP), a DNA end-processing enzyme with 3'-phosphatase and 5'-kinase activities, is involved in multiple DNA repair pathways, including base excision (BER), single-strand break (SSBR), and double-strand break repair (DSBR). However, little is known as to how PNKP functions in such diverse repair processes. Here we report that PNKP is acetylated at K142 (AcK142) by p300 constitutively but at K226 (AcK226) by CBP, only after DSB induction. Co-immunoprecipitation analysis using AcK142 or AcK226 PNKP-specific antibodies showed that AcK142-PNKP associates only with BER/SSBR, and AcK226 PNKP with DSBR proteins. Despite the modest effect of acetylation on PNKP’s enzymatic activity in vitro, cells expressing non-acetylable PNKP (K142R or K226R) accumulated DNA damage in transcribed genes. Intriguingly, in striatal neuronal cells of a Huntington's Disease (HD)-based mouse model, K142, but not K226, was acetylated. This is consistent with the reported degradation of CBP, but not p300, in HD cells. Moreover, transcribed genomes of HD cells progressively accumulated DSBs. Chromatin-immunoprecipitation analysis demonstrated the association of Ac-PNKP with the transcribed genes, consistent with PNKP’s role in transcription-coupled repair. Thus, our findings demonstrate that acetylation at two lysine residues, located in different domains of PNKP, regulates its distinct role in BER/SSBR versus DSBR.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 2470684

Journal Information:: Nucleic Acids Research, Journal Name: Nucleic Acids Research Journal Issue: 5 Vol. 52; ISSN 0305-1048

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: United States

Language:: English

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