Mechanistic studies of the mitotic activation of Mos.
The protein kinase Mos is responsible for the activation of MEK1 and p42 mitogen-activated protein kinase during Xenopus oocyte maturation and during mitosis in Xenopus egg extracts. Here we show that the activation of Mos depends upon the phosphorylation of Ser 3, a residue previously implicated in the regulation of Mos stability; the dephosphorylation of Ser 105, a previously unidentified phosphorylation site conserved in Mos proteins; and the regulated dissociation of Mos from CK2beta. Mutation of Ser 3 to alanine and/or mutation of Ser 105 to glutamate produces a Mos protein that is defective for M-phase activation, as assessed by in vitro kinase assays, and defective for induction of oocyte maturation and maintenance of the spindle assembly checkpoint in extracts. Interestingly, Ser 105 is situated at the beginning of helix alphaC in the N-terminal lobe of the Mos kinase domain. Changes in the orientation of this helix have been previously implicated in the activation of Cdk2 and Src family tyrosine kinases. Our work suggests that Ser 105 dephosphorylation represents a novel mechanism for reorienting helix alphaC.
Yue
Jianbo
J
Stanford University School of Medicine, Department of Molecular Pharmacology, CCSR Room 3155, Stanford, CA 94305-5174, USA. jyue@Stanford.edu
Ferrell
James E
JE
D000818:Animals; D001665:Binding Sites; D047390:Casein Kinase II / Q000378:metabolism; D019926:Cyclin B / Q000378:metabolism; D004789:Enzyme Activation; D005260:Female; D066298:In Vitro Techniques; D008938:Mitosis / Q000502:physiology*; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000166:cytology / Q000201:enzymology; D010766:Phosphorylation; D017231:Proto-Oncogene Proteins c-mos / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012694:Serine / Q000737:chemistry; D014981:Xenopus; D029867:Xenopus Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D019926:Cyclin B; D011994:Recombinant Proteins; D029867:Xenopus Proteins; D012694:Serine; D047390:Casein Kinase II; C495329:Mos protein, Xenopus; D017231:Proto-Oncogene Proteins c-mos