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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),...
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
1997
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2136770/ https://www.ncbi.nlm.nih.gov/pubmed/9281587 |
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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 |
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