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Progression of the mammalian cell cycle is regulated by phosphorylation/dephosphorylation and synthesis/degradation of many key proteins. These events are of utmost importance at the checkpoints, or transition points, of the cell cycle. MAD2 (Mitotic Arrest Deficient) is the human homolog of a yeast and Xenopus protein that is essential for spindle assembly during mitosis. The human hsMAD2 gene encodes a protein of 205 amino acids with a predicted molecular weight of 23.5 kDa. Binding of affinity purified polyclonal antibodies to the MAD2 protein prevents mitosis of HeLa cells. This indicates that, like its invertebrate relatives, MAD2 is necessary for mitosis. Furthermore, MAD2 is localized at the kinetochore of condensed chromosomes during mitosis and cells defective in the mitotic checkpoint have reduced levels of MAD2.
