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Physiological levels of poly(ADP-ribose) during the cell cycle regulate HeLa cell proliferation
Protein targets of polyADP-ribosylation undergo covalent modification with high-molecular-weight, branched poly(ADP-ribose) (PAR) of lengths up to 200 or more ADP-ribose residues derived from NAD(+). PAR polymerase 1 (PARP1) is the most abundant and well-characterized enzyme involved in PAR biosynth...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10009817/ https://www.ncbi.nlm.nih.gov/pubmed/35447104 http://dx.doi.org/10.1016/j.yexcr.2022.113163 |
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author | Yamashita, Sachiko Tanaka, Masakazu Ida, Chieri Kouyama, Kenichi Nakae, Setsu Matsuki, Taisuke Tsuda, Masataka Shirai, Tsuyoshi Kamemura, Kazuo Nishi, Yoshisuke Moss, Joel Miwa, Masanao |
author_facet | Yamashita, Sachiko Tanaka, Masakazu Ida, Chieri Kouyama, Kenichi Nakae, Setsu Matsuki, Taisuke Tsuda, Masataka Shirai, Tsuyoshi Kamemura, Kazuo Nishi, Yoshisuke Moss, Joel Miwa, Masanao |
author_sort | Yamashita, Sachiko |
collection | PubMed |
description | Protein targets of polyADP-ribosylation undergo covalent modification with high-molecular-weight, branched poly(ADP-ribose) (PAR) of lengths up to 200 or more ADP-ribose residues derived from NAD(+). PAR polymerase 1 (PARP1) is the most abundant and well-characterized enzyme involved in PAR biosynthesis. Extensive studies have been carried out to determine how polyADP-ribosylation (PARylation) regulates cell proliferation during cell cycle, with conflicting conclusions. Since significant activation of PARP1 occurs during cell lysis in vitro, we changed the standard method for cell lysis, and using our sensitive ELISA system, quantified without addition of a PAR glycohydrolase inhibitor and clarified that the PAR level is significantly higher in S phase than that in G1. Under normal condition in the absence of exogenous DNA-damaging agent, PAR turns over with a half-life of <40 s; consistent with significant decrease of NAD(+) levels in S phase, which is rescued by PARP inhibitors, in line with the observed rapid turnover of PAR. PARP inhibitors delayed cell cycle in S phase and decreased cell proliferation. Our results underscore the importance of a suitable assay system to measure rapid PAR chain dynamics in living cells and aid our understanding of the function of PARylation during the cell cycle. |
format | Online Article Text |
id | pubmed-10009817 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
record_format | MEDLINE/PubMed |
spelling | pubmed-100098172023-08-01 Physiological levels of poly(ADP-ribose) during the cell cycle regulate HeLa cell proliferation Yamashita, Sachiko Tanaka, Masakazu Ida, Chieri Kouyama, Kenichi Nakae, Setsu Matsuki, Taisuke Tsuda, Masataka Shirai, Tsuyoshi Kamemura, Kazuo Nishi, Yoshisuke Moss, Joel Miwa, Masanao Exp Cell Res Article Protein targets of polyADP-ribosylation undergo covalent modification with high-molecular-weight, branched poly(ADP-ribose) (PAR) of lengths up to 200 or more ADP-ribose residues derived from NAD(+). PAR polymerase 1 (PARP1) is the most abundant and well-characterized enzyme involved in PAR biosynthesis. Extensive studies have been carried out to determine how polyADP-ribosylation (PARylation) regulates cell proliferation during cell cycle, with conflicting conclusions. Since significant activation of PARP1 occurs during cell lysis in vitro, we changed the standard method for cell lysis, and using our sensitive ELISA system, quantified without addition of a PAR glycohydrolase inhibitor and clarified that the PAR level is significantly higher in S phase than that in G1. Under normal condition in the absence of exogenous DNA-damaging agent, PAR turns over with a half-life of <40 s; consistent with significant decrease of NAD(+) levels in S phase, which is rescued by PARP inhibitors, in line with the observed rapid turnover of PAR. PARP inhibitors delayed cell cycle in S phase and decreased cell proliferation. Our results underscore the importance of a suitable assay system to measure rapid PAR chain dynamics in living cells and aid our understanding of the function of PARylation during the cell cycle. 2022-08-01 2022-04-18 /pmc/articles/PMC10009817/ /pubmed/35447104 http://dx.doi.org/10.1016/j.yexcr.2022.113163 Text en https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ). |
spellingShingle | Article Yamashita, Sachiko Tanaka, Masakazu Ida, Chieri Kouyama, Kenichi Nakae, Setsu Matsuki, Taisuke Tsuda, Masataka Shirai, Tsuyoshi Kamemura, Kazuo Nishi, Yoshisuke Moss, Joel Miwa, Masanao Physiological levels of poly(ADP-ribose) during the cell cycle regulate HeLa cell proliferation |
title | Physiological levels of poly(ADP-ribose) during the cell cycle regulate HeLa cell proliferation |
title_full | Physiological levels of poly(ADP-ribose) during the cell cycle regulate HeLa cell proliferation |
title_fullStr | Physiological levels of poly(ADP-ribose) during the cell cycle regulate HeLa cell proliferation |
title_full_unstemmed | Physiological levels of poly(ADP-ribose) during the cell cycle regulate HeLa cell proliferation |
title_short | Physiological levels of poly(ADP-ribose) during the cell cycle regulate HeLa cell proliferation |
title_sort | physiological levels of poly(adp-ribose) during the cell cycle regulate hela cell proliferation |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10009817/ https://www.ncbi.nlm.nih.gov/pubmed/35447104 http://dx.doi.org/10.1016/j.yexcr.2022.113163 |
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