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PMID: 16195348 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Changes in regulatory phosphorylation of Cdc25C Ser287 and Wee1 Ser549 during normal cell cycle progression and checkpoint arrests.

Molecular biology of the cell ·Vol. 16 ·No. 12 ·2005-12-00 ·Pages 5749-60

Stanford JS, Ruderman JV

Abstract

Entry into mitosis is catalyzed by cdc2 kinase. Previous work identified the cdc2-activating phosphatase cdc25C and the cdc2-inhibitory kinase wee1 as targets of the incomplete replication-induced kinase Chk1. Further work led to the model that checkpoint kinases block mitotic entry by inhibiting cdc25C through phosphorylation on Ser287 and activating wee1 through phosphorylation on Ser549. However, almost all conclusions underlying this idea were drawn from work using recombinant proteins. Here, we report that in the early Xenopus egg cell cycles, phosphorylation of endogenous cdc25C Ser287 is normally high during interphase and shows no obvious increase after checkpoint activation. By contrast, endogenous wee1 Ser549 phosphorylation is low during interphase and increases after activation of either the DNA damage or replication checkpoints; this is accompanied by a slight increase in wee1 kinase activity. Blocking mitotic entry by adding the catalytic subunit of PKA also results in increased wee1 Ser549 phosphorylation and maintenance of cdc25C Ser287 phosphorylation. These results argue that in response to checkpoint activation, endogenous wee1 is indeed a critical responder that functions by repressing the cdc2-cdc25C positive feedback loop. Surprisingly, endogenous wee1 Ser549 phosphorylation is highest during mitosis just after the peak of cdc2 activity. Treatments that block inactivation of cdc2 result in further increases in wee1 Ser549 phosphorylation, suggesting a previously unsuspected role for wee1 in mitosis.

MeSH Terms
Animals Cell Cycle/physiology Cell Cycle Proteins/metabolism Cell Nucleus/ultrastructure Cyclic AMP-Dependent Protein Kinases/antagonists & inhibitors,metabolism Female Male Ovum/cytology,ultrastructure Phosphorylation Phosphoserine/metabolism Protein-Tyrosine Kinases/metabolism Spermatozoa/cytology,ultrastructure Xenopus Proteins/metabolism Xenopus laevis cdc25 Phosphatases/metabolism
Chemicals
Cell Cycle Proteins Xenopus Proteins Phosphoserine Wee2 protein, Xenopus Protein-Tyrosine Kinases Cyclic AMP-Dependent Protein Kinases cdc25 Phosphatases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Stanford Jennifer S
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Ruderman Joan V
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e05-06-0541
Published
2005-12-00
Epub
2005-00-29
Pages
5749-60
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1289418
Subset
IM
Grants
NICHD NIH HHS · R01 HD023696 · United States
NICHD NIH HHS · HD-23696 · United States
External Links
PubMed source DOI: 10.1091/mbc.e05-06-0541 PMC Full Text View journal details
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