Regulation of Microtubule Assembly-Disassembly in Mitotic Mammalian Cells: Role of Calcium, Calmodulin and MTOCs

Abstract

Two microtubule arrays exist in cultured mammalian cells during their progression through the cell cycle; the cytoplasmic microtubule complexes (CMTC) of interphase cells (Figure 1) and the mitotic apparatus (MA) of dividing cells (Figure 2). As chromosomes are segregated to opposite poles of the spindle during telophase, the microtubules of the MA are disassembled. During late telophase -G1 phase the tubulin subunits from the spindle are recycled into the microtubules of the CMTC which forms an elaborate network throught the cytoplasm. When cells progress into late G2 -Prophase, the CMTC is disassembled and the tubulin is converted into microtubules of the MA. our research has been aimed at defining the mechanism whereby cells regulate the alternating patterns of microtubule assembly-disassembly during the cell cycle.In one series of experiments, we have investigated the role of calcium in microtubule assembly. Several laboratories have shown that cytoplasmic and spindle microtubules are unstable in the presence of elevated free calcium levels. Using monospecific antibodies and indirect immunofluorescence, we have demonstrated the presence of the ubiquitous calcium-binding protein calmodulin in the mitotic spindle of mammalian cells in vitro (Figure 3).