CDK-mediated phosphorylation of PNKP is required for end-processing of single-strand DNA gaps on Okazaki fragments and genome stability
Polynucleotide kinase phosphatase (PNKP) has enzymatic activities as 3′-phosphatase and 5′-kinase of DNA ends to promote DNA ligation and repair. Here, we show that cyclin-dependent kinases (CDKs) regulate the phosphorylation of threonine 118 (T118) in PNKP. This phosphorylation allows recruitment t...
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| Format: | Article |
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eLife Sciences Publications Ltd
2025-03-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/99217 |
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| author | Kaima Tsukada Rikiya Imamura Tomoko Miyake Kotaro Saikawa Mizuki Saito Naoya Kase Lingyan Fu Masamichi Ishiai Yoshihisa Matsumoto Mikio Shimada |
| author_facet | Kaima Tsukada Rikiya Imamura Tomoko Miyake Kotaro Saikawa Mizuki Saito Naoya Kase Lingyan Fu Masamichi Ishiai Yoshihisa Matsumoto Mikio Shimada |
| author_sort | Kaima Tsukada |
| collection | DOAJ |
| description | Polynucleotide kinase phosphatase (PNKP) has enzymatic activities as 3′-phosphatase and 5′-kinase of DNA ends to promote DNA ligation and repair. Here, we show that cyclin-dependent kinases (CDKs) regulate the phosphorylation of threonine 118 (T118) in PNKP. This phosphorylation allows recruitment to the gapped DNA structure found in single-strand DNA (ssDNA) nicks and/or gaps between Okazaki fragments (OFs) during DNA replication. T118A (alanine)-substituted PNKP-expressing cells exhibited an accumulation of ssDNA gaps in S phase and accelerated replication fork progression. Furthermore, PNKP is involved in poly (ADP-ribose) polymerase 1 (PARP1)-dependent replication gap filling as part of a backup pathway in the absence of OFs ligation. Altogether, our data suggest that CDK-mediated PNKP phosphorylation at T118 is important for its recruitment to ssDNA gaps to proceed with OFs ligation and its backup repairs via the gap-filling pathway to maintain genome stability. |
| format | Article |
| id | doaj-art-6dbffeee1d30479293b4586d7b6eb6e0 |
| institution | Kabale University |
| issn | 2050-084X |
| language | English |
| publishDate | 2025-03-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj-art-6dbffeee1d30479293b4586d7b6eb6e02025-08-20T03:42:02ZengeLife Sciences Publications LtdeLife2050-084X2025-03-011410.7554/eLife.99217CDK-mediated phosphorylation of PNKP is required for end-processing of single-strand DNA gaps on Okazaki fragments and genome stabilityKaima Tsukada0https://orcid.org/0000-0002-6725-9514Rikiya Imamura1Tomoko Miyake2https://orcid.org/0000-0003-3389-4007Kotaro Saikawa3Mizuki Saito4Naoya Kase5Lingyan Fu6Masamichi Ishiai7https://orcid.org/0000-0003-4313-9945Yoshihisa Matsumoto8https://orcid.org/0000-0002-0758-290XMikio Shimada9https://orcid.org/0000-0003-1980-9187Laboratory for Zero-Carbon Energy, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, Japan; Center for Chromosome Stability, Department of Cellular and Molecular Medicine, University of Copenhagen, Copenhagen, DenmarkLaboratory for Zero-Carbon Energy, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, JapanLaboratory for Zero-Carbon Energy, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, JapanLaboratory for Zero-Carbon Energy, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, JapanLaboratory for Zero-Carbon Energy, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, JapanLaboratory for Zero-Carbon Energy, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, JapanLaboratory for Zero-Carbon Energy, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, JapanNational Cancer Center Research Institute, Tokyo, JapanLaboratory for Zero-Carbon Energy, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, JapanLaboratory for Zero-Carbon Energy, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, JapanPolynucleotide kinase phosphatase (PNKP) has enzymatic activities as 3′-phosphatase and 5′-kinase of DNA ends to promote DNA ligation and repair. Here, we show that cyclin-dependent kinases (CDKs) regulate the phosphorylation of threonine 118 (T118) in PNKP. This phosphorylation allows recruitment to the gapped DNA structure found in single-strand DNA (ssDNA) nicks and/or gaps between Okazaki fragments (OFs) during DNA replication. T118A (alanine)-substituted PNKP-expressing cells exhibited an accumulation of ssDNA gaps in S phase and accelerated replication fork progression. Furthermore, PNKP is involved in poly (ADP-ribose) polymerase 1 (PARP1)-dependent replication gap filling as part of a backup pathway in the absence of OFs ligation. Altogether, our data suggest that CDK-mediated PNKP phosphorylation at T118 is important for its recruitment to ssDNA gaps to proceed with OFs ligation and its backup repairs via the gap-filling pathway to maintain genome stability.https://elifesciences.org/articles/99217PNKPDNA replicationOkazaki fragmentDNA repair |
| spellingShingle | Kaima Tsukada Rikiya Imamura Tomoko Miyake Kotaro Saikawa Mizuki Saito Naoya Kase Lingyan Fu Masamichi Ishiai Yoshihisa Matsumoto Mikio Shimada CDK-mediated phosphorylation of PNKP is required for end-processing of single-strand DNA gaps on Okazaki fragments and genome stability eLife PNKP DNA replication Okazaki fragment DNA repair |
| title | CDK-mediated phosphorylation of PNKP is required for end-processing of single-strand DNA gaps on Okazaki fragments and genome stability |
| title_full | CDK-mediated phosphorylation of PNKP is required for end-processing of single-strand DNA gaps on Okazaki fragments and genome stability |
| title_fullStr | CDK-mediated phosphorylation of PNKP is required for end-processing of single-strand DNA gaps on Okazaki fragments and genome stability |
| title_full_unstemmed | CDK-mediated phosphorylation of PNKP is required for end-processing of single-strand DNA gaps on Okazaki fragments and genome stability |
| title_short | CDK-mediated phosphorylation of PNKP is required for end-processing of single-strand DNA gaps on Okazaki fragments and genome stability |
| title_sort | cdk mediated phosphorylation of pnkp is required for end processing of single strand dna gaps on okazaki fragments and genome stability |
| topic | PNKP DNA replication Okazaki fragment DNA repair |
| url | https://elifesciences.org/articles/99217 |
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