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PMID: 12119357 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A role for regulated binding of p150(Glued) to microtubule plus ends in organelle transport.

The Journal of cell biology ·Vol. 158 ·No. 2 ·2002-07-22 ·Pages 305-19

Vaughan PS, Miura P, Henderson M, Byrne B, Vaughan KT

Abstract

A subset of microtubule-associated proteins, including cytoplasmic linker protein (CLIP)-170, dynactin, EB1, adenomatous polyposis coli, cytoplasmic dynein, CLASPs, and LIS-1, has been shown recently to target to the plus ends of microtubules. The mechanisms and functions of this binding specificity are not understood, although a role in encouraging microtubule elongation has been proposed. To extend previous work on the role of dynactin in organelle transport, we analyzed p150(Glued) by live-cell imaging. Time-lapse analysis of p150(Glued) revealed targeting to the plus ends of growing microtubules, requiring the NH2-terminal cytoskeleton-associated protein-glycine rich domain, but not EB1 or CLIP-170. Effectors of protein kinase A modulated microtubule binding and suggested p150(Glued) phosphorylation as a factor in plus-end binding specificity. Using a phosphosensitive monoclonal antibody, we mapped the site of p150(Glued) phosphorylation to Ser-19. In vivo and in vitro analysis of phosphorylation site mutants revealed that p150(Glued) phosphorylation mediates dynamic binding to microtubules. To address the function of dynamic binding, we imaged GFP-p150(Glued) during the dynein-dependent transport of Golgi membranes. Live-cell analysis revealed a transient interaction between Golgi membranes and GFP-p150(Glued)-labeled microtubules just prior to transport, implicating microtubules and dynactin in a search-capture mechanism for minus-end-directed organelles.

MeSH Terms
Animals Binding Sites/genetics Biological Transport COS Cells Dynactin Complex Dyneins/metabolism Golgi Apparatus/metabolism Microtubule-Associated Proteins/chemistry,metabolism Microtubules/chemistry,metabolism,ultrastructure Mutagenesis, Site-Directed Phosphorylation Protein Binding
Chemicals
Dynactin Complex Microtubule-Associated Proteins Dyneins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Vaughan Patricia S
Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA.
Miura Pedro
Henderson Matthew
Byrne Belinda
Vaughan Kevin T
References (46)
46 references, click to expand
  1. Interaction of the p62 subunit of dynactin with Arp1 and the cortical actin cytoskeleton.
    Curr Biol. 1999 Dec 16-30;9(24):1497-500 PMID: 10607597
  2. Evidence for a role of CLIP-170 in the establishment of metaphase chromosome alignment.
    J Cell Biol. 1998 May 18;141(4):849-62 PMID: 9585405
  3. Adenomatous polyposis coli (APC) protein moves along microtubules and concentrates at their growing ends in epithelial cells.
    J Cell Biol. 2000 Feb 7;148(3):505-18 PMID: 10662776
  4. Microtubule interactions with the cell cortex causing nuclear movements in Saccharomyces cerevisiae.
    J Cell Biol. 2000 May 15;149(4):863-74 PMID: 10811827
  5. Functional analysis of CLIP-115 and its binding to microtubules.
    J Cell Sci. 2000 Jun;113 ( Pt 12):2285-97 PMID: 10825300
  6. Dynamics of cytoplasmic dynein in living cells and the effect of a mutation in the dynactin complex actin-related protein Arp1.
    Curr Biol. 2000 May 18;10(10):603-6 PMID: 10837229
  7. The dynamic behavior of the APC-binding protein EB1 on the distal ends of microtubules.
    Curr Biol. 2000 Jul 13;10(14):865-8 PMID: 10899006
  8. Dynein, dynactin, and kinesin II's interaction with microtubules is regulated during bidirectional organelle transport.
    J Cell Biol. 2000 Oct 2;151(1):155-66 PMID: 11018061
  9. A role for the lissencephaly gene LIS1 in mitosis and cytoplasmic dynein function.
    Nat Cell Biol. 2000 Nov;2(11):784-91 PMID: 11056532
  10. Immunolocalization of cytoplasmic dynein and dynactin subunits in cultured macrophages: enrichment on early endocytic organelles.
    J Cell Sci. 2001 Jan;114(Pt 1):229-240 PMID: 11112706
  11. Search, capture and signal: games microtubules and centrosomes play.
    J Cell Sci. 2001 Jan;114(Pt 2):247-55 PMID: 11148127
  12. Clasps are CLIP-115 and -170 associating proteins involved in the regional regulation of microtubule dynamics in motile fibroblasts.
    Cell. 2001 Mar 23;104(6):923-35 PMID: 11290329
  13. Cytoplasmic dynein intermediate chain phosphorylation regulates binding to dynactin.
    J Biol Chem. 2001 Jul 13;276(28):26171-9 PMID: 11340075
  14. Role of dynein, dynactin, and CLIP-170 interactions in LIS1 kinetochore function.
    J Cell Biol. 2002 Mar 18;156(6):959-68 PMID: 11889140
  15. LIS1, CLIP-170's key to the dynein/dynactin pathway.
    Mol Cell Biol. 2002 May;22(9):3089-102 PMID: 11940666
  16. The polarity and stability of microtubule capture by the kinetochore.
    J Cell Biol. 1988 Jan;106(1):151-9 PMID: 3339086
  17. Direct observation of microtubule dynamics in living cells.
    Nature. 1988 Apr 21;332(6166):724-6 PMID: 3357537
  18. Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin A suggests an ER recycling pathway.
    Cell. 1990 Mar 9;60(5):821-36 PMID: 2178778
  19. Kinetochores capture astral microtubules during chromosome attachment to the mitotic spindle: direct visualization in live newt lung cells.
    J Cell Biol. 1990 Sep;111(3):1039-45 PMID: 2391359
  20. Homology of a 150K cytoplasmic dynein-associated polypeptide with the Drosophila gene Glued.
    Nature. 1991 Jun 13;351(6327):579-83 PMID: 1828535
  21. Binding of pp170 to microtubules is regulated by phosphorylation.
    J Biol Chem. 1991 Sep 15;266(26):17597-605 PMID: 1680130
  22. Two activators of microtubule-based vesicle transport.
    J Cell Biol. 1991 Dec;115(5):1309-18 PMID: 1835460
  23. Dynactin, a conserved, ubiquitously expressed component of an activator of vesicle motility mediated by cytoplasmic dynein.
    J Cell Biol. 1991 Dec;115(6):1639-50 PMID: 1836789
  24. Cytoplasmic dynein participates in the centrosomal localization of the Golgi complex.
    J Cell Biol. 1992 Sep;118(6):1333-45 PMID: 1387874
  25. CLIP-170 links endocytic vesicles to microtubules.
    Cell. 1992 Sep 18;70(6):887-900 PMID: 1356075
  26. Characterization of a 50-kDa polypeptide in cytoplasmic dynein preparations reveals a complex with p150GLUED and a novel actin.
    J Biol Chem. 1993 Jul 15;268(20):15318-23 PMID: 8325901
  27. The p150Glued component of the dynactin complex binds to both microtubules and the actin-related protein centractin (Arp-1).
    Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1634-8 PMID: 7878030
  28. Modulation of microtubule dynamic instability in vivo by brain microtubule associated proteins.
    J Cell Sci. 1995 Apr;108 ( Pt 4):1679-89 PMID: 7615685
  29. Affinity chromatography demonstrates a direct binding between cytoplasmic dynein and the dynactin complex.
    J Biol Chem. 1995 Dec 1;270(48):28806-11 PMID: 7499404
  30. Cytoplasmic dynein binds dynactin through a direct interaction between the intermediate chains and p150Glued.
    J Cell Biol. 1995 Dec;131(6 Pt 1):1507-16 PMID: 8522607
  31. Fluorescent speckle microscopy, a method to visualize the dynamics of protein assemblies in living cells.
    Curr Biol. 1998 Nov 5;8(22):1227-30 PMID: 9811609
  32. EB1, a protein which interacts with the APC tumour suppressor, is associated with the microtubule cytoskeleton throughout the cell cycle.
    Oncogene. 1998 Dec 31;17(26):3471-7 PMID: 10030671
  33. CLIP-170 highlights growing microtubule ends in vivo.
    Cell. 1999 Feb 19;96(4):517-27 PMID: 10052454
  34. Colocalization of cytoplasmic dynein with dynactin and CLIP-170 at microtubule distal ends.
    J Cell Sci. 1999 May;112 ( Pt 10):1437-47 PMID: 10212138
  35. The APC-associated protein EB1 associates with components of the dynactin complex and cytoplasmic dynein intermediate chain.
    Curr Biol. 1999 Apr 22;9(8):425-8 PMID: 10226031
  36. Role of dynactin in endocytic traffic: effects of dynamitin overexpression and colocalization with CLIP-170.
    Mol Biol Cell. 1999 Dec;10(12):4107-20 PMID: 10588646
  37. Properties of the kinetochore in vitro. II. Microtubule capture and ATP-dependent translocation.
    J Cell Biol. 1985 Sep;101(3):766-77 PMID: 4030894
  38. The adenomatous polyposis coli tumor suppressor protein localizes to plasma membrane sites involved in active cell migration.
    J Cell Biol. 1996 Jul;134(1):165-79 PMID: 8698812
  39. Centractin (ARP1) associates with spectrin revealing a potential mechanism to link dynactin to intracellular organelles.
    J Cell Biol. 1996 Dec;135(6 Pt 2):1815-29 PMID: 8991093
  40. Dynactin phosphorylation is modulated in response to cellular effectors.
    Biochem Biophys Res Commun. 1997 Mar 27;232(3):810-6 PMID: 9126359
  41. Partitioning of the Golgi apparatus during mitosis in living HeLa cells.
    J Cell Biol. 1997 Jun 16;137(6):1211-28 PMID: 9182657
  42. Microtubules orient the mitotic spindle in yeast through dynein-dependent interactions with the cell cortex.
    J Cell Biol. 1997 Aug 11;138(3):629-41 PMID: 9245791
  43. ER-to-Golgi transport visualized in living cells.
    Nature. 1997 Sep 4;389(6646):81-5 PMID: 9288971
  44. Overexpression of the dynamitin (p50) subunit of the dynactin complex disrupts dynein-dependent maintenance of membrane organelle distribution.
    J Cell Biol. 1997 Oct 20;139(2):469-84 PMID: 9334349
  45. Microtubule polymerization dynamics.
    Annu Rev Cell Dev Biol. 1997;13:83-117 PMID: 9442869
  46. Dynactin increases the processivity of the cytoplasmic dynein motor.
    Nat Cell Biol. 2000 Jan;2(1):20-4 PMID: 10620802
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200201029
Published
2002-07-22
Epub
2002-00-15
Pages
305-19
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2173134
Subset
IM
Grants
NIGMS NIH HHS · R01 GM060560 · United States
NIGMS NIH HHS · GM 60560 · United States
External Links
PubMed source DOI: 10.1083/jcb.200201029 PMC Full Text View journal details
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