Proteins specifically associated with the microtubules of the mammalian mitotic spindle

Abstract

The molecular species that determine the unique structure and functions of the microtubules in the mitotic spindle are not known. We describe the results of two new approaches to the molecular structure of the spindle. Both approaches rely on detergent-extracted preparations of synchronized populations of cells metabolically labeled with 35S-methionine or 32P-phosphate. In these preparations, the original cellular microtubules are preserved. The microtubule components can be released from the detergent-extracted preparations by selective depolymerization with calcium ions. Alternatively, the microtubules can be stabilized by taxol, freed of chromatin by digestion with DNAase and freed of the surrounding cage of intermediate filaments by further extraction at low ionic strength. Gel electrophoresis of each of these preparations of mitotic microtubules demonstrates that they contain microtubule-associated proteins that we have previously shown to be present in interphase microtubules. They also contain a protein of 150,000 daltons, which is the first mitosis-specific microtubule-associated protein identified in mammalian cells.