Home LiteratureArticle Details
PMID: 20935625 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Direct dynamin-actin interactions regulate the actin cytoskeleton.

The EMBO journal ·Vol. 29 ·No. 21 ·2010-11-03 ·Pages 3593-606

Gu C, Yaddanapudi S, Weins A, Osborn T, Reiser J, Pollak M, Hartwig J, Sever S

Abstract

The large GTPase dynamin assembles into higher order structures that are thought to promote endocytosis. Dynamin also regulates the actin cytoskeleton through an unknown, GTPase-dependent mechanism. Here, we identify a highly conserved site in dynamin that binds directly to actin filaments and aligns them into bundles. Point mutations in the actin-binding domain cause aberrant membrane ruffling and defective actin stress fibre formation in cells. Short actin filaments promote dynamin assembly into higher order structures, which in turn efficiently release the actin-capping protein (CP) gelsolin from barbed actin ends in vitro, allowing for elongation of actin filaments. Together, our results support a model in which assembled dynamin, generated through interactions with short actin filaments, promotes actin polymerization via displacement of actin-CPs.

MeSH Terms
Actin Cytoskeleton/metabolism Actins/metabolism Amino Acid Sequence Animals Cells, Cultured Cytoskeleton/metabolism Dynamins/antagonists & inhibitors,genetics,metabolism Endocytosis/physiology Gelsolin/metabolism HeLa Cells Humans Mice Molecular Sequence Data Muscle, Skeletal/cytology,metabolism Podocytes/metabolism Protein Binding RNA, Small Interfering/pharmacology Rabbits Sequence Homology, Amino Acid Stress Fibers/metabolism
Chemicals
Actins Gelsolin RNA, Small Interfering Dynamins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Gu Changkyu
Nephrology Division, Department of Medicine, Harvard Medical School and Massachusetts General Hospital, Charlestown, MA 02129, USA.
Yaddanapudi Suma
Weins Astrid
Osborn Teresia
Reiser Jochen
Pollak Martin
Hartwig John
Sever Sanja
References (55)
55 references, click to expand
  1. The molecular and functional phenotype of glomerular podocytes reveals key features of contractile smooth muscle cells.
    Am J Physiol Renal Physiol. 2008 Oct;295(4):F959-70 PMID: 18684887
  2. The actin binding site of thymosin beta 4 mapped by mutational analysis.
    EMBO J. 1996 Jan 15;15(2):201-10 PMID: 8617195
  3. Dynamin at actin tails.
    Proc Natl Acad Sci U S A. 2002 Jan 8;99(1):161-6 PMID: 11782545
  4. Ca2+ control of actin filament length. Effects of macrophage gelsolin on actin polymerization.
    J Biol Chem. 1981 Sep 25;256(18):9693-7 PMID: 6270098
  5. Dynamin as a mover and pincher during cell migration and invasion.
    J Cell Sci. 2006 May 1;119(Pt 9):1683-90 PMID: 16636070
  6. Rapid purification of native dynamin I and colorimetric GTPase assay.
    Methods Enzymol. 2005;404:556-69 PMID: 16413300
  7. Cellular motility driven by assembly and disassembly of actin filaments.
    Cell. 2003 Feb 21;112(4):453-65 PMID: 12600310
  8. Role of formins in actin assembly: nucleation and barbed-end association.
    Science. 2002 Jul 26;297(5581):612-5 PMID: 12052901
  9. Impairment of dynamin's GAP domain stimulates receptor-mediated endocytosis.
    Nature. 1999 Apr 8;398(6727):481-6 PMID: 10206643
  10. A role for gelsolin in stress fiber-dependent cell contraction.
    Exp Cell Res. 1999 Jul 10;250(1):155-67 PMID: 10388529
  11. Three-dimensional reconstruction of dynamin in the constricted state.
    Nat Cell Biol. 2001 Oct;3(10):922-6 PMID: 11584275
  12. Reorganization of the actin cytoskeleton in the protruding lamellae of human fibroblasts.
    Cell Motil Cytoskeleton. 2001 Sep;50(1):13-32 PMID: 11746669
  13. Expression, purification, and functional assays for self-association of dynamin-1.
    Methods Enzymol. 2001;329:447-57 PMID: 11210565
  14. SNX9 couples actin assembly to phosphoinositide signals and is required for membrane remodeling during endocytosis.
    Dev Cell. 2007 Jul;13(1):43-56 PMID: 17609109
  15. Cofilin changes the twist of F-actin: implications for actin filament dynamics and cellular function.
    J Cell Biol. 1997 Aug 25;138(4):771-81 PMID: 9265645
  16. Rearrangements of the cytoskeleton and cell contacts induce process formation during differentiation of conditionally immortalized mouse podocyte cell lines.
    Exp Cell Res. 1997 Oct 10;236(1):248-58 PMID: 9344605
  17. Synaptopodin orchestrates actin organization and cell motility via regulation of RhoA signalling.
    Nat Cell Biol. 2006 May;8(5):485-91 PMID: 16622418
  18. Direct observation of the uncapping of capping protein-capped actin filaments by CARMIL homology domain 3.
    J Biol Chem. 2010 Jan 22;285(4):2707-20 PMID: 19926785
  19. Induction of mutant dynamin specifically blocks endocytic coated vesicle formation.
    J Cell Biol. 1994 Nov;127(4):915-34 PMID: 7962076
  20. Purification of muscle actin.
    Methods Enzymol. 1982;85 Pt B:164-81 PMID: 7121269
  21. The large GTPase dynamin regulates actin comet formation and movement in living cells.
    Proc Natl Acad Sci U S A. 2002 Jan 8;99(1):167-72 PMID: 11782546
  22. Proteolytic processing of dynamin by cytoplasmic cathepsin L is a mechanism for proteinuric kidney disease.
    J Clin Invest. 2007 Aug;117(8):2095-104 PMID: 17671649
  23. Gelsolin, a protein that caps the barbed ends and severs actin filaments, enhances the actin-based motility of Listeria monocytogenes in host cells.
    Infect Immun. 1998 Aug;66(8):3775-82 PMID: 9673261
  24. Evidence for a gelsolin-rich, labile F-actin pool in human polymorphonuclear leukocytes.
    Cell Motil Cytoskeleton. 1992;21(1):25-37 PMID: 1311641
  25. Dissecting dynamin's role in clathrin-mediated endocytosis.
    Biochem Soc Trans. 2009 Oct;37(Pt 5):1022-6 PMID: 19754444
  26. Dynamin2 GTPase and cortactin remodel actin filaments.
    J Biol Chem. 2009 Sep 4;284(36):23995-4005 PMID: 19605363
  27. Regulating actin dynamics at membranes: a focus on dynamin.
    Traffic. 2004 Jul;5(7):463-9 PMID: 15180823
  28. Formin proteins: purification and measurement of effects on actin assembly.
    Methods Enzymol. 2006;406:215-34 PMID: 16472660
  29. Control of actin polymerization in live and permeabilized fibroblasts.
    J Cell Biol. 1991 Aug;114(3):503-13 PMID: 1860882
  30. Dynamin2 and cortactin regulate actin assembly and filament organization.
    Curr Biol. 2002 Oct 29;12(21):1852-7 PMID: 12419186
  31. Robust colorimetric assays for dynamin's basal and stimulated GTPase activities.
    Methods Enzymol. 2005;404:490-503 PMID: 16413294
  32. Mutational and Biological Analysis of alpha-actinin-4 in focal segmental glomerulosclerosis.
    J Am Soc Nephrol. 2005 Dec;16(12):3694-701 PMID: 16251236
  33. Dynamin self-assembly stimulates its GTPase activity.
    J Biol Chem. 1996 Sep 13;271(37):22310-4 PMID: 8798389
  34. A continuous, regenerative coupled GTPase assay for dynamin-related proteins.
    Methods Enzymol. 2005;404:611-9 PMID: 16413304
  35. Dynamic (re)organization of the podocyte actin cytoskeleton in the nephrotic syndrome.
    Pediatr Nephrol. 2004 Feb;19(2):130-7 PMID: 14673634
  36. Effects of cytochalasin and phalloidin on actin.
    J Cell Biol. 1987 Oct;105(4):1473-8 PMID: 3312229
  37. Actin nucleation and elongation factors: mechanisms and interplay.
    Curr Opin Cell Biol. 2009 Feb;21(1):28-37 PMID: 19168341
  38. Actin, a central player in cell shape and movement.
    Science. 2009 Nov 27;326(5957):1208-12 PMID: 19965462
  39. EGF stimulates an increase in actin nucleation and filament number at the leading edge of the lamellipod in mammary adenocarcinoma cells.
    J Cell Sci. 1998 Jan;111 ( Pt 2):199-211 PMID: 9405304
  40. Single molecule kinetic analysis of actin filament capping. Polyphosphoinositides do not dissociate capping proteins.
    J Biol Chem. 2007 Sep 21;282(38):28014-24 PMID: 17656356
  41. The variable twist of actin and its modulation by actin-binding proteins.
    J Cell Biol. 1987 Apr;104(4):1005-17 PMID: 3558475
  42. Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape.
    J Cell Biol. 2000 Oct 2;151(1):187-98 PMID: 11018064
  43. The plasma and cytoplasmic forms of human gelsolin differ in disulfide structure.
    Biochemistry. 1996 Jul 30;35(30):9700-9 PMID: 8703941
  44. Domain structure and intramolecular regulation of dynamin GTPase.
    EMBO J. 1997 Nov 17;16(22):6676-83 PMID: 9362482
  45. The dynamin superfamily: universal membrane tubulation and fission molecules?
    Nat Rev Mol Cell Biol. 2004 Feb;5(2):133-47 PMID: 15040446
  46. Alternative splicing of the actin binding domain of human cortactin affects cell migration.
    J Biol Chem. 2003 Nov 14;278(46):45672-9 PMID: 12952985
  47. Coordinated regulation of platelet actin filament barbed ends by gelsolin and capping protein.
    J Cell Biol. 1996 Jul;134(2):389-99 PMID: 8707824
  48. Relationship between Arp2/3 complex and the barbed ends of actin filaments at the leading edge of carcinoma cells after epidermal growth factor stimulation.
    J Cell Biol. 1999 Apr 19;145(2):331-45 PMID: 10209028
  49. Dynamin at the actin-membrane interface.
    Curr Opin Cell Biol. 2003 Feb;15(1):31-9 PMID: 12517701
  50. Ubiquitously expressed dynamin-II has a higher intrinsic GTPase activity and a greater propensity for self-assembly than neuronal dynamin-I.
    Mol Biol Cell. 1997 Dec;8(12):2553-62 PMID: 9398675
  51. Kinetic mechanism of end-to-end annealing of actin filaments.
    J Mol Biol. 2001 Sep 28;312(4):721-30 PMID: 11575927
  52. Dynamin self-assembles into rings suggesting a mechanism for coated vesicle budding.
    Nature. 1995 Mar 9;374(6518):190-2 PMID: 7877694
  53. Gelsolin deficiency blocks podosome assembly and produces increased bone mass and strength.
    J Cell Biol. 2000 Feb 21;148(4):665-78 PMID: 10684249
  54. Dynamin forms a Src kinase-sensitive complex with Cbl and regulates podosomes and osteoclast activity.
    Mol Biol Cell. 2005 Jul;16(7):3301-13 PMID: 15872089
  55. Stabilization and the cytoplasmic ground substance in detergent-opened cells and a structural and biochemical analysis of its composition.
    Proc Natl Acad Sci U S A. 1981 Jul;78(7):4329-33 PMID: 6945586
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.1038/emboj.2010.249
Published
2010-11-03
Epub
2010-00-08
Pages
3593-606
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2982766
Subset
IM
Grants
NIDDK NIH HHS · R01 DK059588 · United States
NIDDK NIH HHS · R01 DK059588-10 · United States
NIDDK NIH HHS · R01 DK064787 · United States
NIDDK NIH HHS · R01 DK64787 · United States
Corrections
CommentIn
PMID: 21045865
External Links
PubMed source DOI: 10.1038/emboj.2010.249 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2