Sil phosphorylation in a Pin1 binding domain affects the duration of the spindle checkpoint.
SIL is an immediate-early gene that is essential for embryonic development and is implicated in T-cell leukemia-associated translocations. We now show that the Sil protein is hyperphosphorylated during mitosis or in cells blocked at prometaphase by microtubule inhibitors. Cell cycle-dependent phosphorylation of Sil is required for its interaction with Pin1, ... a regulator of mitosis. Point mutation of the seven (S/T)P sites between amino acids 567 and 760 reduces mitotic phosphorylation of Sil, Pin1 binding, and spindle checkpoint duration. When a phosphorylation site mutant Sil is stably expressed, the duration of the spindle checkpoint is shortened in cells challenged with taxol or nocodazole, and the cells revert to a G2-like state. This event is associated with the downregulation of the kinase activity of the Cdc2/cyclin B1 complex and the dephosphorylation of the threonine 161 on the Cdc2 subunit. Sil downregulation by plasmid-mediated RNA interference limited the ability of cells to activate the spindle checkpoint and correlated with a reduction of Cdc2/cyclin B1 activity and phosphorylation on T161 on the Cdc2 subunit. These data suggest that a critical region of Sil is required to mediate the presentation of Cdc2 activity during spindle checkpoint arrest.
Mesh Terms:
Binding Sites, Cell Cycle, DNA, HeLa Cells, Humans, Intracellular Signaling Peptides and Proteins, Mitosis, NIMA-Interacting Peptidylprolyl Isomerase, Oncogene Proteins, Fusion, Peptidylprolyl Isomerase, Phosphorylation, Protein Binding, Protein Structure, Tertiary, RNA Interference, Spindle Apparatus, Time Factors
Binding Sites, Cell Cycle, DNA, HeLa Cells, Humans, Intracellular Signaling Peptides and Proteins, Mitosis, NIMA-Interacting Peptidylprolyl Isomerase, Oncogene Proteins, Fusion, Peptidylprolyl Isomerase, Phosphorylation, Protein Binding, Protein Structure, Tertiary, RNA Interference, Spindle Apparatus, Time Factors
Mol Cell Biol
Date: Aug. 01, 2005
PubMed ID: 16024801
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