Preparation of microtubules from rat liver and testis: cytoplasmic dynein is a major microtubule associated protein.
A microtubule associated protein from brain tissue (MAP 1C), has been found to possess many properties in common with ciliary and flagellar dyneins (Paschal et al.:J. Cell Biol. 105:1273-1282, 1987). However, this protein, now designated as cytoplasmic dynein, exhibited several properties which distinguish it from axonemal forms of the enzyme. We have investigated these characteristics further in a study of cytoplasmic dyneins from non-neuronal tissues. Rat liver and testis in particular were found to contain high levels of cytoplasmic dynein. The yield of dynein from testis was over 70 micrograms/g of tissue, making this the best source of cytoplasmic dynein of all tissues so far examined. The characterization of dynein from these sources has confirmed and extended our previous observations concerning the unique properties of cytoplasmic dynein. Activation of liver and testis dynein occurred at low (less than 1 mg/ml) tubulin concentration. Polypeptides identified as subunits of brain cytoplasmic dynein (74, 59, 57, 55, and 53 kDa) were present in liver and testis preparations. In addition, polypeptides at 150 and 45 kDa were found to copurify with the non-neuronal dyneins. The liver and testis enzyme hydrolyzed pyrimidine nucleotides at rates up to 12.5 times faster than ATP, though the relative affinity of cytoplasmic dynein for CTP was much lower (Km = 1.0 mM) than that for ATP. The properties of the testis enzyme were consistent with its identification as a cytoplasmic dynein rather than a sperm axonemal precursor. These data indicate that cytoplasmic dyneins may be widespread in distribution and that they share certain biochemical properties unique from those of axonemal dyneins. These characteristics are consistent with the proposal that cytoplasmic dynein plays a universal role in retrograde organelle motility.
Collins
C A
CA
Cell Biology Group, Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts.
Vallee
R B
RB
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D003593:Cytoplasm / Q000201:enzymology; D004398:Dyneins / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D008099:Liver / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000201:enzymology* / Q000378:metabolism; D009708:Nucleotidases / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013737:Testis / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure
D008869:Microtubule-Associated Proteins; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D009708:Nucleotidases; D000251:Adenosine Triphosphatases; D004398:Dyneins