SPIN, A substrate in the MAP kinase pathway in mouse oocytes

The newly cloned gene Spin encodes a 30-kDa protein, a well-defined abundant molecule found in mouse oocytes and early embryos. This protein SPIN undergoes metaphase-specific phosphorylation and binds to the spindle. To understand the role of SPIN in oocyte meiosis, oocytes were treated with drugs that affect the cell cycle by activating or inactivating specific kinases. The posttranslation modification of SPIn in the treated oocytes was then investigated by one- and two-dimensional gel electrophoresis. Modification of SPIn is inhibited by treatment with 6-dimethyl-aminopurine (DMAP), suggesting that SPIN is phosphorylated by a serine-threonine kinase. Furthermore, SPIN from cycloheximide-treated oocytes that lack detectable MAP kinase activity is only partially phosphrylated, indicating that SPIN may be phosphorylated by the MOS/MAP kinase pathway. To confirm this observation, SPIN was analyzed in Mos-null mutant mice lacking MAP kinase activity. Normal posttranslational modification of SPIN was analyzed in Mos-null mutant mice lacking MAP kinase activity pathway and further that this phosphorylation of SPIN may be essential for its interaction with the spindle.