DIRECTIONAL INSTABILITY OF MICROTUBULE TRANSPORT IN THE PRESENCE OF KINESIN AND DYNEIN, 2 OPPOSITE POLARITY MOTOR PROTEINS

被引:109
作者
VALE, RD [1 ]
MALIK, F [1 ]
BROWN, D [1 ]
机构
[1] UNIV CALIF SAN FRANCISCO, DEPT BIOCHEM, SAN FRANCISCO, CA 94143 USA
关键词
D O I
10.1083/jcb.119.6.1589
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Kinesin and dynein are motor proteins that move in opposite directions along microtubules. In this study, we examine the consequences of having kinesin and dynein (ciliary outer arm or cytoplasmic) bound to glass surfaces interacting with the same microtubule in vitro. Although one might expect a balance of opposing forces to produce little or no net movement, we find instead that microtubules move unidirectionally for several microns (corresponding to hundreds of ATPase cycles by a motor) but continually switch between kinesin-directed and dynein-directed transport. The velocities in the plus-end (0.2-0.3 mum/s) and minus-end (3.5-4 mum/s) directions were approximately half those produced by kinesin (0.5 mum/s) and ciliary dynein (6.7 mum/s) alone, indicating that the motors not contributing to movement can interact with and impose a drag upon the microtubule. By comparing two dyneins with different duty ratios (percentage of time spent in a strongly bound state during the ATPase cycle) and varying the nucleotide conditions, we show that the microtubule attachment times of the two opposing motors as well as their relative numbers determine which motor predominates in this assay. Together, these findings are consistent with a model in which kinesin-induced movement of a microtubule induces a negative strain in attached dyneins which causes them to dissociate before entering a force-generating state (and vice versa); reversals in the direction of transport may require the temporary dissociation of the transporting motor from the microtubule. The bidirectional movements described here are also remarkably similar to the back-and-forth movements of chromosomes during mitosis and membrane vesicles in fibroblasts. These results suggest that the underlying mechanical properties of motor proteins, at least in part, may be responsible for reversals in microtubule-based transport observed in cells.
引用
收藏
页码:1589 / 1596
页数:8
相关论文
共 45 条
[1]   FAST AXONAL-TRANSPORT IN SQUID GIANT-AXON [J].
ALLEN, RD ;
METUZALS, J ;
TASAKI, I ;
BRADY, ST ;
GILBERT, SP .
SCIENCE, 1982, 218 (4577) :1127-1129
[3]   BEAD MOVEMENT BY SINGLE KINESIN MOLECULES STUDIED WITH OPTICAL TWEEZERS [J].
BLOCK, SM ;
GOLDSTEIN, LSB ;
SCHNAPP, BJ .
NATURE, 1990, 348 (6299) :348-352
[5]   TRACTION FORCE ON A KINETOCHORE AT METAPHASE ACTS AS A LINEAR FUNCTION OF KINETOCHORE FIBER LENGTH [J].
HAYS, TS ;
WISE, D ;
SALMON, ED .
JOURNAL OF CELL BIOLOGY, 1982, 93 (02) :374-382
[6]   A TIME-LAPSE VIDEO IMAGE INTENSIFICATION ANALYSIS OF CYTOPLASMIC ORGANELLE MOVEMENTS DURING ENDOSOME TRANSLOCATION [J].
HERMAN, B ;
ALBERTINI, DF .
JOURNAL OF CELL BIOLOGY, 1984, 98 (02) :565-576
[7]   MOVEMENT OF MICROTUBULES BY SINGLE KINESIN MOLECULES [J].
HOWARD, J ;
HUDSPETH, AJ ;
VALE, RD .
NATURE, 1989, 342 (6246) :154-158
[8]   MUSCLE STRUCTURE AND THEORIES OF CONTRACTION [J].
HUXLEY, AF .
PROGRESS IN BIOPHYSICS & MOLECULAR BIOLOGY, 1957, 7 :255-&
[9]   NOTE SUGGESTING THAT CROSS-BRIDGE ATTACHMENT DURING MUSCLE-CONTRACTION MAY TAKE PLACE IN 2 STAGES [J].
HUXLEY, AF .
PROCEEDINGS OF THE ROYAL SOCIETY SERIES B-BIOLOGICAL SCIENCES, 1973, 183 (1070) :83-86
[10]  
HYMAN A, 1990, METHOD ENZYMOL, V196, P303