Xenopus interphase and mitotic microtubule-associated proteins differentially suppress microtubule dynamics in vitro

Based on observations of microtubule dynamics in Xenopus extracts and in vivo, it has been assumed that the pool of interphase microtubule-associated proteins (MAPs) are more potent microtubule stabilizers than their mitotic counterparts. The aim of this study was to test that assumption, and two questions were addressed here. First, are there differences in the composition of interphase and mitotic MAPs? Second, do interphase MAPs more potently promote microtubule assembly than mitotic MAPs? Biochemical purification from Xenopus egg extracts shows that the composition of interphase and mitotic MAPs is similar. XMAP215, XMAP230, and XMAP310, which are the three characterized Xenopus MAPs, show decreased microtubule binding in mitotic extracts, and mitotic MAPs are slightly more phosphorylated than interphase MAPs. Bulk polymerization and time-lapse video microscopy show that microtubules polymerized two times faster in the presence of total interphase MAPs compared with total mitotic MAPs. Interphase but not mitotic MAPs strongly promoted microtubule nucleation in solution. Video microscopy showed that microtubules never underwent catastrophes in the presence of either MAP fraction. It is proposed that the increase in microtubule dynamics at the onset of mitosis results from phosphorylation dependent decreased microtubule stabilization by MAPs, allowing destabilizing factors to increase the catastrophe frequency and dismantle the interphase microtubule network.