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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...

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Autores principales: Yamashita, Sachiko, Tanaka, Masakazu, Ida, Chieri, Kouyama, Kenichi, Nakae, Setsu, Matsuki, Taisuke, Tsuda, Masataka, Shirai, Tsuyoshi, Kamemura, Kazuo, Nishi, Yoshisuke, Moss, Joel, Miwa, Masanao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2022
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.
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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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