Brain dynein crossbridges microtubules into bundles.
Cytoplasmic dynein was purified from pig brain, using a modified version of published procedures, in order to study its interaction with microtubules. Since the preparation produces ATP-dependent sliding of taxol-stabilized purified microtubules over glass and runs on SDS-containing gels as a major band exceeding 300,000 Mr plus a medium chain band at about 75,000 Mr, it is assumed to be identical to the mammalian brain dynein (MAP 1C) purified by Vallee and colleagues. When viewed by electron microscopy in negative stain, individual particles show two distinct configurations. Some are clearly similar to the two-headed bouquet structure already shown for MAP 1C. A larger number of molecules in the present preparation appear to have two heads fused together, forming a dimeric globular particle with two separate tails. They are referred to as phiparticles, because of their resemblance to the greek letter phi. A model for the structural relationship between the two molecular forms is presented. The stems of two associated dynein subunits may separate beyond the base, to form a bouquet, or they may remain fused to form the larger tail of a phi-particle. The smaller tail probably represents a combined pair of features equivalent to the 'stalks' shown to emanate from axonemal dynein heads by Goodenough and colleagues. Both tails of a phi-particle can bind to microtubules, even in the presence of ATP, and cause microtubule bundling. These results suggest a complete structural homology between axonemal and cytoplasmic dynein.
Amos
L A
LA
MRC Laboratory of Molecular Biology, Cambridge, UK.
D000251:Adenosine Triphosphatases; D000818:Animals; D001921:Brain / Q000201:enzymology / Q000648:ultrastructure*; D004398:Dyneins / Q000302:isolation & purification / Q000648:ultrastructure*; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008870:Microtubules / Q000201:enzymology / Q000648:ultrastructure*; D008961:Models, Structural; D008970:Molecular Weight; D013552:Swine
D046911:Macromolecular Substances; D000251:Adenosine Triphosphatases; D004398:Dyneins