Thromb Haemost 1997; 78(02): 910-918
DOI: 10.1055/s-0038-1657651
Rapid Communication
Schattauer GmbH Stuttgart

Activation of Human Platelets Causes Post-translational Modifications to Cytoplasmic Dynein

Stephen W Rothwell
The Department of Hematology and Vascular Biology, Walter Reed Army Institute of Research, Washington, D.C., USA
,
Valerie S Calvert
The Department of Hematology and Vascular Biology, Walter Reed Army Institute of Research, Washington, D.C., USA
› Author Affiliations
Further Information

Publication History

Received 06 1996

Accepted after resubmission 26 March 1997

Publication Date:
12 July 2018 (online)

Summary

In our studies of human platelets we have detected the presence of the molecular motors kinesin and dynein. Dynein is present at a concentration (0.8 μg/g tissue) that is approximately 1/3 the concentration reported for neuronal tissue. Immunofluorescence microscopy of resting platelets shows that, while platelet microtubules are arranged in coiled hoops forming the marginal band in the cortical region of the platelet, dynein is distributed in a pattern of punctate staining throughout the cytoplasm of the platelets. Fractionation of unactivated platelets shows that dynein partitions to the soluble fraction. Stimulation of platelets with thrombin, ADP or epinephrine causes a partial translocation of dynein from the soluble fraction to the particulate fraction with thrombin being the most efficient agent at promoting this shift. Dynein intermediate chain recovered in the soluble fraction of disrupted platelets following activation displays a transient, time-dependent phosphorylation. In contrast, dynein intermediate chain recovered in the particulate fraction shows decreased phosphorylation. These results indicate that human platelets contain a complex microtubule-based system of motor proteins that is an integral part of the physiological changes occurring during platelet activation.

 
  • References

  • 1 White JG. Ultrastructural analysis of platelet contractile apparatus. Meth Enzymol 1992; 215: 109-127
  • 2 Fox JEB. The platelet cytoskeleton. Thromb Haemost 1993; 70: 884-893
  • 3 Hartwig JH. Mechanism of actin rearrangements mediating platelet activation. J Cell Biol 1992; 118: 1421-1442
  • 4 White JG. Arrangements of actin filaments in the cytoskeleton of human platelets. Am J Pathol 1984; 117: 207-217
  • 5 Debus E, Weber K, Osborn M. The cytoskeleton of blood platelets viewed by immunofluorescence microscopy. Eur J Cell Biol 1981; 24: 45-52
  • 6 Beckerle MC, Miller DE, Bertagnolli ME. Activation-dependent redistribution of the adhesion plaque protein, talin, in intact human platelets. J Cell Biol 1989; 109: 3333-3346
  • 7 Fox JEB, Phillips DR. Role of phosphorylation in mediating the association of myosin with the cytoskeletonal structures of human platelets. J Biol Chem 1982; 257: 4120-4126
  • 8 Kenney DM, Chao FC. Ionophore-induced disassembly of blood platelet microtubules: effect of cyclic AMP and indomethacin. J Cell Physiol 1980; 103: 289-298
  • 9 Steiner M, Ikeda Y. Quantitative assessment of polymerized and depolymerized platelets microtubules. Changes caused by aggregating agents. J Clin Invest 1979; 63: 443-452
  • 10 White JG, Sauk JJ. Microtubule coils in spread blood platelets. Blood 1984; 64: 470-478
  • 11 Cohen I, Gerrard JM, White JG. Ultrastructure of clots during isometric contraction. J Cell Biol 1982; 93: 775-787
  • 12 Yano Y, Sakon M, Kambayashi J, Kawasaki T, Senda T, Tanaka K, Yamada F, Shibata N. Cytoskeletal reorganization of human platelets induced by the protein phosphatase ½ A inhibitors okadaic acid and calyculin A. BiochemJ 1995; 307: 439-449
  • 13 Schroer TA, Steuer ER, Sheetz MP. Cytoplasmic dynein is a minus end- directed motor for membranous organelles. Cell 1989; 56: 937-946
  • 14 Hymen AA, Mitchison TJ. Two different microtubule-based motor activities with opposite polarities in kinetochores. Nature 1991; 351: 206-211
  • 15 Steuer ER, Wordeman L, Schroer TA, Sheetz MP. Localization of cytoplasmic dynein to mitotic spindles and kinetochores. Nature 1990; 345: 266-268
  • 16 Lippincott-Schwartz J. Membrane cycling between the ER and Golgi apparatus and its role in biosynthetic transport. Subcell Biochem 1993; 21: 095-119
  • 17 Lippincott-Schwartz J, Cole NB, Marotta A, Conrad PA, Bloom GS. Kinesin is the motor for microtubule-mediated Golgi-to-ER membrane traffic [published erratum appears in J Cell Biol 1995 Mar; 128: following 988]. J Cell Biol 1995; 128: 293-306
  • 18 Feiguin F, Ferreira A, Kosik KS, Caceres A. Kinesin-mediated organelle translocation revealed by specific cellular manipulations. J Cell Biol 1994; 127: 1021-1039
  • 19 Aniento F, Emans N, Griffiths G, Gruenberg J. Cytoplasmic dynein-depen- dent vesicular transport from early to late endosomes. J Cell Biol 1993; 123: 1373-1387
  • 20 Holzbaur ELF, Vallee R. Dyneins: Molecular structure and cellular function. Annu Rev Cell Biol 1994; 10: 339-372
  • 21 Stenberg PE, McDonald TP, Jackson CW. Disruption of microtubules in vivo by vincristo induces large membrane complexes and other cytoplasmic abnormalities in megakaryocytes and platelets of normal rats like those of human and Wistar Furth rat hereditary macrothrombocytopenias. J Cell Physiol 1995; 162: 086-102
  • 22 Berry S, Dawicki DD, Agarwal KC, Steiner M. The role of microtubules in platelet secretory release. Biochim Biophys Acta 1989; 1012: 46-56
  • 23 White JG. Effects of colchicine and vinca alkaloids on human platelets. Influence on platelet microtubules and contractile function. Am J Pathol 1968; 53: 281-291
  • 24 White JG, Rao GH. Influence of a microtubule stabilizing agent on platelet structural physiology. Am J Pathol 1983; 112: 207-217
  • 25 Handagama PJ, George JN, Shuman MA, McEver RP, Bainton DF. Incorporation of a circulating protein into megakaryocyte and platelet granules. Proc Nat Acad Sci (USA) 1987; 84: 861-865
  • 26 Belitser N, Anischuk M, Veklich Y, Pozdnjakova T, Gorkun O. Fibrinogen internalization by ADP-stimulated blood platelets. Ultrastructural studies with fibrinogen-colloidal gold probes. Thromb Res 1993; 69: 413-424
  • 27 Handagama P, Scarborough RM, Shuman MA, Bainton DF. Endocytosis of fibrinogen into megakaryocyte and platelet alpha-granules is mediated by alpha lib beta 3 (glycoprotein Ilb-IIIa). Blood 1993; 82: 135-138
  • 28 Suzuki M, Kawakatsu T, Nagata H, Hamamoto K, Iwata K, Ohga S, Nomura S, Kokawa T, Yasunaga K. Effects of injected antibody against the platelet glycoprotein Ilb/HIa complex on monkey platelet fibrinogen. Thromb Haemost 1992; 67: 578-581
  • 29 Wenzel-Drake JD, Frelinger IIIAL, Dieter MG, Lam SC-T. Arg-Gly-Aspdependent occupancy of GP Ilb/IIIa by applagin: Evidence for internalization and cycling of a platelet integrin. Blood 1993; 81: 62-69
  • 30 Brass LF, Ahuja M, Belmonte E, Blanchard N, Pizarro S, Tarver A, Hoxie JA. Thrombin receptors: turning them off after turning them on. Semin Hematol 1994; 31: 251-260
  • 31 Escolar G, White JG. The platelet open canalicular system: a final common pathway. Blood Cells 1991; 17: 467-485
  • 32 Murphy DB, Wallis KL. Isolation of microtubule protein from chicken erythrocytes and determination of the critical concentration for tubulin polymerization in vitro and in vivo. J Biol Chem 1983; 258: 8357-8364
  • 33 Towbin H, Staehlin T, Gordon J. Electrophorectic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedures and some applications. Proc Natl Acad Sci (USA) 1979; 76: 4350-4354
  • 34 Rothwell SW, Deal CC, Pinto J, Wright DG. Affinity purification and sub- cellular localization of kinesin in human neutrophils. J Leukoc Biol 1993; 53: 372-380
  • 35 Nasr A, Satir P. Alloaffmity Filtration: A General Approach to the Purification of Dynein and Dynein-like Molecules. Anal Biochem 1985; 151: 097-108
  • 36 Paschal BM, Shpetner HS, Vallee RB. MAP 1C is a microtubule-activated ATPase which translocates microtubules in vitro and dynein-like properties. J Cell Biol 1987; 105: 1273-1282
  • 37 Cooper JA. Effects of cytochalasin and phalloidin on actin. J Cell Biol 1987; 105: 1473-1478
  • 38 Schnapp BJ, Reese TS. Dynein is the motor for retrograde axonal transport of organelles. Proc Natl Acad Sci (USA) 1989; 86: 1548-1552
  • 39 Pfister KK, Salata MW, Dillman JF, Vaughan KT, Vallee RB, Torre E, Lye RJ. Differential expression and phosphorylation of the 74 kDa intermediate chains of cytoplasmic dynein in cultured neurons and glia. J Biol Chem 1996; 271: 1687-1694
  • 40 Johnson LN, Barford D. Electrostatic effects in the control of glycogen phosphorylase by phosphorylation. Protein Science 1994; 03: 1726-1730
  • 41 Lin SX, Ferro KL, Collins CA. Cytoplasmic dynein undergoes intracellular redistribution concomitant with phosphorylation of the heavy chain in response to serum starvation and okadaic acid. J Cell Biol 1994; 127: 1009-1019
  • 42 Lacey ML, Haimo LT. Cytoplasmic dynein binds to phospholipid vesicles. Cell Motil Cytoskeleton 1994; 28: 205-212