Cargando…

A Dual-Specificity Phosphatase Cdc25B Is an Unstable Protein and Triggers p34(cdc2)/Cyclin B Activation in Hamster BHK21 Cells Arrested with Hydroxyurea

By incubating at 30°C in the presence of an energy source, p34(cdc2)/cyclin B was activated in the extract prepared from a temperature-sensitive mutant, tsBN2, which prematurely enters mitosis at 40°C, the nonpermissive temperature (Nishimoto, T., E. Eilen, and C. Basilico. 1978. Cell. 15:475–483),...

Descripción completa

Detalles Bibliográficos
Autores principales: Nishijima, Hitoshi, Nishitani, Hideo, Seki, Takashi, Nishimoto, Takeharu
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1997
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2136770/
https://www.ncbi.nlm.nih.gov/pubmed/9281587
_version_ 1782143197193961472
author Nishijima, Hitoshi
Nishitani, Hideo
Seki, Takashi
Nishimoto, Takeharu
author_facet Nishijima, Hitoshi
Nishitani, Hideo
Seki, Takashi
Nishimoto, Takeharu
author_sort Nishijima, Hitoshi
collection PubMed
description By incubating at 30°C in the presence of an energy source, p34(cdc2)/cyclin B was activated in the extract prepared from a temperature-sensitive mutant, tsBN2, which prematurely enters mitosis at 40°C, the nonpermissive temperature (Nishimoto, T., E. Eilen, and C. Basilico. 1978. Cell. 15:475–483), and wild-type cells of the hamster BHK21 cell line arrested in S phase, without protein synthesis. Such an in vitro activation of p34(cdc2)/cyclin B, however, did not occur in the extract prepared from cells pretreated with protein synthesis inhibitor cycloheximide, although this extract still retained the ability to inhibit p34(cdc2)/cyclin B activation. When tsBN2 cells arrested in S phase were incubated at 40°C in the presence of cycloheximide, Cdc25B, but not Cdc25A and C, among a family of dual-specificity phosphatases, Cdc25, was lost coincidentally with the lack of the activation of p34(cdc2)/cyclin B. Consistently, the immunodepletion of Cdc25B from the extract inhibited the activation of p34(cdc2)/cyclin B. Cdc25B was found to be unstable (half-life < 30 min). Cdc25B, but not Cdc25C, immunoprecipitated from the extract directly activated the p34(cdc2)/cyclin B of cycloheximide-treated cells as well as that of nontreated cells, although Cdc25C immunoprecipitated from the extract of mitotic cells activated the p34(cdc2)/cyclin B within the extract of cycloheximide-treated cells. Our data suggest that Cdc25B made an initial activation of p34(cdc2)/cyclin B, which initiates mitosis through the activation of Cdc25C.
format Text
id pubmed-2136770
institution National Center for Biotechnology Information
language English
publishDate 1997
publisher The Rockefeller University Press
record_format MEDLINE/PubMed
spelling pubmed-21367702008-05-01 A Dual-Specificity Phosphatase Cdc25B Is an Unstable Protein and Triggers p34(cdc2)/Cyclin B Activation in Hamster BHK21 Cells Arrested with Hydroxyurea Nishijima, Hitoshi Nishitani, Hideo Seki, Takashi Nishimoto, Takeharu J Cell Biol Article By incubating at 30°C in the presence of an energy source, p34(cdc2)/cyclin B was activated in the extract prepared from a temperature-sensitive mutant, tsBN2, which prematurely enters mitosis at 40°C, the nonpermissive temperature (Nishimoto, T., E. Eilen, and C. Basilico. 1978. Cell. 15:475–483), and wild-type cells of the hamster BHK21 cell line arrested in S phase, without protein synthesis. Such an in vitro activation of p34(cdc2)/cyclin B, however, did not occur in the extract prepared from cells pretreated with protein synthesis inhibitor cycloheximide, although this extract still retained the ability to inhibit p34(cdc2)/cyclin B activation. When tsBN2 cells arrested in S phase were incubated at 40°C in the presence of cycloheximide, Cdc25B, but not Cdc25A and C, among a family of dual-specificity phosphatases, Cdc25, was lost coincidentally with the lack of the activation of p34(cdc2)/cyclin B. Consistently, the immunodepletion of Cdc25B from the extract inhibited the activation of p34(cdc2)/cyclin B. Cdc25B was found to be unstable (half-life < 30 min). Cdc25B, but not Cdc25C, immunoprecipitated from the extract directly activated the p34(cdc2)/cyclin B of cycloheximide-treated cells as well as that of nontreated cells, although Cdc25C immunoprecipitated from the extract of mitotic cells activated the p34(cdc2)/cyclin B within the extract of cycloheximide-treated cells. Our data suggest that Cdc25B made an initial activation of p34(cdc2)/cyclin B, which initiates mitosis through the activation of Cdc25C. The Rockefeller University Press 1997-09-08 /pmc/articles/PMC2136770/ /pubmed/9281587 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Article
Nishijima, Hitoshi
Nishitani, Hideo
Seki, Takashi
Nishimoto, Takeharu
A Dual-Specificity Phosphatase Cdc25B Is an Unstable Protein and Triggers p34(cdc2)/Cyclin B Activation in Hamster BHK21 Cells Arrested with Hydroxyurea
title A Dual-Specificity Phosphatase Cdc25B Is an Unstable Protein and Triggers p34(cdc2)/Cyclin B Activation in Hamster BHK21 Cells Arrested with Hydroxyurea
title_full A Dual-Specificity Phosphatase Cdc25B Is an Unstable Protein and Triggers p34(cdc2)/Cyclin B Activation in Hamster BHK21 Cells Arrested with Hydroxyurea
title_fullStr A Dual-Specificity Phosphatase Cdc25B Is an Unstable Protein and Triggers p34(cdc2)/Cyclin B Activation in Hamster BHK21 Cells Arrested with Hydroxyurea
title_full_unstemmed A Dual-Specificity Phosphatase Cdc25B Is an Unstable Protein and Triggers p34(cdc2)/Cyclin B Activation in Hamster BHK21 Cells Arrested with Hydroxyurea
title_short A Dual-Specificity Phosphatase Cdc25B Is an Unstable Protein and Triggers p34(cdc2)/Cyclin B Activation in Hamster BHK21 Cells Arrested with Hydroxyurea
title_sort dual-specificity phosphatase cdc25b is an unstable protein and triggers p34(cdc2)/cyclin b activation in hamster bhk21 cells arrested with hydroxyurea
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2136770/
https://www.ncbi.nlm.nih.gov/pubmed/9281587
work_keys_str_mv AT nishijimahitoshi adualspecificityphosphatasecdc25bisanunstableproteinandtriggersp34cdc2cyclinbactivationinhamsterbhk21cellsarrestedwithhydroxyurea
AT nishitanihideo adualspecificityphosphatasecdc25bisanunstableproteinandtriggersp34cdc2cyclinbactivationinhamsterbhk21cellsarrestedwithhydroxyurea
AT sekitakashi adualspecificityphosphatasecdc25bisanunstableproteinandtriggersp34cdc2cyclinbactivationinhamsterbhk21cellsarrestedwithhydroxyurea
AT nishimototakeharu adualspecificityphosphatasecdc25bisanunstableproteinandtriggersp34cdc2cyclinbactivationinhamsterbhk21cellsarrestedwithhydroxyurea
AT nishijimahitoshi dualspecificityphosphatasecdc25bisanunstableproteinandtriggersp34cdc2cyclinbactivationinhamsterbhk21cellsarrestedwithhydroxyurea
AT nishitanihideo dualspecificityphosphatasecdc25bisanunstableproteinandtriggersp34cdc2cyclinbactivationinhamsterbhk21cellsarrestedwithhydroxyurea
AT sekitakashi dualspecificityphosphatasecdc25bisanunstableproteinandtriggersp34cdc2cyclinbactivationinhamsterbhk21cellsarrestedwithhydroxyurea
AT nishimototakeharu dualspecificityphosphatasecdc25bisanunstableproteinandtriggersp34cdc2cyclinbactivationinhamsterbhk21cellsarrestedwithhydroxyurea