Analysis of the Dynein-dynactin Interaction in Vitro and in Vivo

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2003 Oct 21

PMID 14565986

Citations 104

Authors

Stephen J King

Christa L Brown

Kerstin C Maier

Nicholas J Quintyne

Trina A Schroer

Affiliations

Soon will be listed here.

Abstract

Cytoplasmic dynein and dynactin are megadalton-sized multisubunit molecules that function together as a cytoskeletal motor. In the present study, we explore the mechanism of dynein-dynactin binding in vitro and then extend our findings to an in vivo context. Solution binding assays were used to define binding domains in the dynein intermediate chain (IC) and dynactin p150Glued subunit. Transient overexpression of a series of fragments of the dynein IC was used to determine the importance of this subunit for dynein function in mammalian tissue culture cells. Our results suggest that a functional dynein-dynactin interaction is required for proper microtubule organization and for the transport and localization of centrosomal components and endomembrane compartments. The dynein IC fragments have different effects on endomembrane localization, suggesting that different endomembranes may bind dynein via distinct mechanisms.

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References

Boylan K, Serr M, HAYS T . A molecular genetic analysis of the interaction between the cytoplasmic dynein intermediate chain and the glued (dynactin) complex. Mol Biol Cell. 2000; 11(11):3791-803. PMC: 15037. DOI: 10.1091/mbc.11.11.3791. View

Wittmann T, Hyman T . Recombinant p50/dynamitin as a tool to examine the role of dynactin in intracellular processes. Methods Cell Biol. 1999; 61:137-43. DOI: 10.1016/s0091-679x(08)61978-0. View

Habermann A, Schroer T, Griffiths G, Burkhardt J . Immunolocalization of cytoplasmic dynein and dynactin subunits in cultured macrophages: enrichment on early endocytic organelles. J Cell Sci. 2000; 114(Pt 1):229-240. DOI: 10.1242/jcs.114.1.229. View

MURESAN V, Stankewich M, Steffen W, Morrow J, Holzbaur E, Schnapp B . Dynactin-dependent, dynein-driven vesicle transport in the absence of membrane proteins: a role for spectrin and acidic phospholipids. Mol Cell. 2001; 7(1):173-83. DOI: 10.1016/s1097-2765(01)00165-4. View

Lo K, Naisbitt S, Fan J, Sheng M, Zhang M . The 8-kDa dynein light chain binds to its targets via a conserved (K/R)XTQT motif. J Biol Chem. 2001; 276(17):14059-66. DOI: 10.1074/jbc.M010320200. View

Mok Y, Lo K, Zhang M . Structure of Tctex-1 and its interaction with cytoplasmic dynein intermediate chain. J Biol Chem. 2001; 276(17):14067-74. DOI: 10.1074/jbc.M011358200. View

Lane J, Vergnolle M, Woodman P, Allan V . Apoptotic cleavage of cytoplasmic dynein intermediate chain and p150(Glued) stops dynein-dependent membrane motility. J Cell Biol. 2001; 153(7):1415-26. PMC: 2150727. DOI: 10.1083/jcb.153.7.1415. View

Tai A, Chuang J, Sung C . Cytoplasmic dynein regulation by subunit heterogeneity and its role in apical transport. J Cell Biol. 2001; 153(7):1499-509. PMC: 2150720. DOI: 10.1083/jcb.153.7.1499. View

Vaughan P, Leszyk J, Vaughan K . Cytoplasmic dynein intermediate chain phosphorylation regulates binding to dynactin. J Biol Chem. 2001; 276(28):26171-9. DOI: 10.1074/jbc.M102649200. View

10.

Holleran E, Ligon L, Tokito M, Stankewich M, Morrow J, Holzbaur E . beta III spectrin binds to the Arp1 subunit of dynactin. J Biol Chem. 2001; 276(39):36598-605. DOI: 10.1074/jbc.M104838200. View

11.

Makokha M, Hare M, Li M, Hays T, Barbar E . Interactions of cytoplasmic dynein light chains Tctex-1 and LC8 with the intermediate chain IC74. Biochemistry. 2002; 41(13):4302-11. DOI: 10.1021/bi011970h. View

12.

King S, Bonilla M, Rodgers M, Schroer T . Subunit organization in cytoplasmic dynein subcomplexes. Protein Sci. 2002; 11(5):1239-50. PMC: 2373548. DOI: 10.1110/ps.2520102. View

13.

Susalka S, Nikulina K, Salata M, Vaughan P, King S, Vaughan K . The roadblock light chain binds a novel region of the cytoplasmic Dynein intermediate chain. J Biol Chem. 2002; 277(36):32939-46. DOI: 10.1074/jbc.M205510200. View

14.

Quintyne N, Schroer T . Distinct cell cycle-dependent roles for dynactin and dynein at centrosomes. J Cell Biol. 2002; 159(2):245-54. PMC: 2173046. DOI: 10.1083/jcb.200203089. View

15.

Deacon S, Serpinskaya A, Vaughan P, Lopez Fanarraga M, Vernos I, Vaughan K . Dynactin is required for bidirectional organelle transport. J Cell Biol. 2003; 160(3):297-301. PMC: 2172679. DOI: 10.1083/jcb.200210066. View

16.

Eckley D, Schroer T . Interactions between the evolutionarily conserved, actin-related protein, Arp11, actin, and Arp1. Mol Biol Cell. 2003; 14(7):2645-54. PMC: 165665. DOI: 10.1091/mbc.e03-01-0049. View

17.

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

18.

Towbin H, Staehelin T, Gordon J . Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979; 76(9):4350-4. PMC: 411572. DOI: 10.1073/pnas.76.9.4350. View

19.

Salisbury J, Baron A, Coling D, Martindale V, Sanders M . Calcium-modulated contractile proteins associated with the eucaryotic centrosome. Cell Motil Cytoskeleton. 1986; 6(2):193-7. DOI: 10.1002/cm.970060218. View

20.

Schweizer A, Fransen J, Bachi T, Ginsel L, Hauri H . Identification, by a monoclonal antibody, of a 53-kD protein associated with a tubulo-vesicular compartment at the cis-side of the Golgi apparatus. J Cell Biol. 1988; 107(5):1643-53. PMC: 2115344. DOI: 10.1083/jcb.107.5.1643. View