THROMBIN, EPIDERMAL GROWTH-FACTOR, AND PHORBOL-MYRISTATE ACETATE STIMULATE TUBULIN POLYMERIZATION IN QUIESCENT CELLS - A POTENTIAL LINK TO MITOGENESIS

被引:12
作者
BALL, RL
ALBRECHT, T
THOMPSON, WC
JAMES, O
CARNEY, DH [1 ]
机构
[1] UNIV TEXAS, MED BRANCH, DEPT HUMAN BIOL CHEM & GENET, GALVESTON, TX 77550 USA
[2] UNIV TEXAS, MED BRANCH, DEPT MICROBIOL, GALVESTON, TX 77550 USA
来源
CELL MOTILITY AND THE CYTOSKELETON | 1992年 / 23卷 / 04期
关键词
GROWTH FACTORS; PHORBOL; 12-MYRISTATE; 13-ACETATE; MICROTUBULE-TUBULIN EQUILIBRIUM; INITIATION OF DNA SYNTHESIS;
D O I
10.1002/cm.970230406
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Previous studies suggest that alterations in the microtubule (MT)-tubulin equilibrium during G0/G1 affect mitogenesis. To determine the effect of growth factors on the MT-tubulin equilibrium, we developed a radioactive monoclonal antibody binding assay (Ball et al.: J. Cell. Biol. 103:1033-1041, 1986). With this assay, H-3-Ab 1-1. 1 binding to cytoskeletons in confluent populations of cultured cells is proportional to the number of tubulin subunits polymerized into MTs. We now show that purified alpha-thrombin increase H-3-Ab 1-1.1 binding to cytoskeletons of serum-arrested mouse embryo (ME) fibroblasts from 1.5- to 3-fold. This stimulation is dose-dependent and correlates with concentrations of thrombin required for initiation of DNA synthesis. Other mitogenic factors, epidermal growth factor (EGF) and phorbol 12-myristate 13-acetate (PMA), also stimulate MT polymerization. Addition of colchicine (0.3 muM) eight hours after growth factor addition, blocks stimulation of H-3-thymidine incorporation by thrombin, EGF, or PMA, suggesting that tubulin polymerization or subsequent events triggered by MT polymerization are required for cells to enter a proliferative cycle. Consistent with models for autoregulation of tubulin synthesis, thrombin, EGF, and PMA all increase tubulin synthesis 9 to 15 hr after growth factor addition, raising the possibility that the decrease in free tubulin and subsequent stimulation of tubulin synthesis is linked to progression of cells into a proliferative cycle. Colchicine addition to these cells also stimulates DNA synthesis, but colchicine-stimulated cells enter S phase 6 to 8 hr later than those stimulated by growth factors. This delayed stimulation may be related to the time required for degradation of tubulin-colchicine complexes below a critical level. These data suggest that regulation of cell proliferation may be linked to increased MT polymerization and the resulting decrease in free tubulin pools.
引用
收藏
页码:265 / 278
页数:14
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