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PMID: 12975351 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Molecular requirements for actin-based lamella formation in Drosophila S2 cells.

The Journal of cell biology ·Vol. 162 ·No. 6 ·2003-09-15 ·Pages 1079-88

Rogers SL, Wiedemann U, Stuurman N, Vale RD

Abstract

Cell migration occurs through the protrusion of the actin-enriched lamella. Here, we investigated the effects of RNAi depletion of approximately 90 proteins implicated in actin function on lamella formation in Drosophila S2 cells. Similar to in vitro reconstitution studies of actin-based Listeria movement, we find that lamellae formation requires a relatively small set of proteins that participate in actin nucleation (Arp2/3 and SCAR), barbed end capping (capping protein), filament depolymerization (cofilin and Aip1), and actin monomer binding (profilin and cyclase-associated protein). Lamellae are initiated by parallel and partially redundant signaling pathways involving Rac GTPases and the adaptor protein Nck, which stimulate SCAR, an Arp2/3 activator. We also show that RNAi of three proteins (kette, Abi, and Sra-1) known to copurify with and inhibit SCAR in vitro leads to SCAR degradation, revealing a novel function of this protein complex in SCAR stability. Our results have identified an essential set of proteins involved in actin dynamics during lamella formation in Drosophila S2 cells.

MeSH Terms
Actin Cytoskeleton/metabolism,ultrastructure Actin Depolymerizing Factors Actin-Related Protein 2 Actins/antagonists & inhibitors,deficiency,genetics,metabolism Adaptor Proteins, Signal Transducing Animals Caenorhabditis elegans Proteins/antagonists & inhibitors,genetics Carrier Proteins/antagonists & inhibitors,genetics,metabolism Cell Cycle Proteins/antagonists & inhibitors,genetics,metabolism Cell Line Cell Movement/physiology Contractile Proteins Cytoskeletal Proteins/antagonists & inhibitors,genetics,metabolism Destrin Drosophila Proteins/antagonists & inhibitors,deficiency,genetics,metabolism Drosophila melanogaster Eukaryotic Cells/cytology,metabolism Microfilament Proteins/antagonists & inhibitors,deficiency,genetics Profilins Pseudopodia/metabolism,ultrastructure RNA Interference rac GTP-Binding Proteins/metabolism
Chemicals
Abi protein, Drosophila Actin Depolymerizing Factors Actin-Related Protein 2 Actins Adaptor Proteins, Signal Transducing CAP1 protein, human Caenorhabditis elegans Proteins Capt protein, Drosophila Carrier Proteins Cell Cycle Proteins Contractile Proteins Cytoskeletal Proteins Destrin Drosophila Proteins Microfilament Proteins Profilins SCAR protein, Drosophila Sra-1 protein, Drosophila actin interacting protein 1 arx-2 protein, C elegans chic protein, Drosophila rac GTP-Binding Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Rogers Stephen L
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94107, USA.
Wiedemann Ursula
Stuurman Nico
Vale Ronald D
References (39)
39 references, click to expand
  1. Control of actin assembly and disassembly at filament ends.
    Curr Opin Cell Biol. 2000 Feb;12(1):97-103 PMID: 10679358
  2. Reconstitution of actin-based motility of Listeria and Shigella using pure proteins.
    Nature. 1999 Oct 7;401(6753):613-6 PMID: 10524632
  3. Use of double-stranded RNA interference in Drosophila cell lines to dissect signal transduction pathways.
    Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6499-503 PMID: 10823906
  4. act up controls actin polymerization to alter cell shape and restrict Hedgehog signaling in the Drosophila eye disc.
    Cell. 2000 Apr 28;101(3):271-81 PMID: 10847682
  5. Repulsive axon guidance: Abelson and Enabled play opposing roles downstream of the roundabout receptor.
    Cell. 2000 Jun 23;101(7):703-15 PMID: 10892742
  6. Molecular mechanisms controlling actin filament dynamics in nonmuscle cells.
    Annu Rev Biophys Biomol Struct. 2000;29:545-76 PMID: 10940259
  7. A cyclase-associated protein regulates actin and cell polarity during Drosophila oogenesis and in yeast.
    Curr Biol. 2000 Aug 24;10(16):964-73 PMID: 10985383
  8. Cell motility: proline-rich proteins promote protrusions.
    Trends Cell Biol. 2001 Jan;11(1):38-46 PMID: 11146297
  9. Wasp, the Drosophila Wiskott-Aldrich syndrome gene homologue, is required for cell fate decisions mediated by Notch signaling.
    J Cell Biol. 2001 Jan 8;152(1):1-13 PMID: 11149916
  10. Reconstitution of human Arp2/3 complex reveals critical roles of individual subunits in complex structure and activity.
    Mol Cell. 2001 Nov;8(5):1041-52 PMID: 11741539
  11. Control of actin reorganization by Slingshot, a family of phosphatases that dephosphorylate ADF/cofilin.
    Cell. 2002 Jan 25;108(2):233-46 PMID: 11832213
  12. SCAR is a primary regulator of Arp2/3-dependent morphological events in Drosophila.
    J Cell Biol. 2002 Feb 18;156(4):689-701 PMID: 11854309
  13. The lamellipodium: where motility begins.
    Trends Cell Biol. 2002 Mar;12(3):112-20 PMID: 11859023
  14. Rac function and regulation during Drosophila development.
    Nature. 2002 Mar 28;416(6879):438-42 PMID: 11919634
  15. Antagonism between Ena/VASP proteins and actin filament capping regulates fibroblast motility.
    Cell. 2002 May 17;109(4):509-21 PMID: 12086607
  16. Molecular dissection of cytokinesis by RNA interference in Drosophila cultured cells.
    Mol Biol Cell. 2002 Jul;13(7):2448-60 PMID: 12134082
  17. Mechanism of regulation of WAVE1-induced actin nucleation by Rac1 and Nck.
    Nature. 2002 Aug 15;418(6899):790-3 PMID: 12181570
  18. Drosophila EB1 is important for proper assembly, dynamics, and positioning of the mitotic spindle.
    J Cell Biol. 2002 Sep 2;158(5):873-84 PMID: 12213835
  19. Cellular control of actin nucleation.
    Annu Rev Cell Dev Biol. 2002;18:247-88 PMID: 12142287
  20. Rho GTPases in cell biology.
    Nature. 2002 Dec 12;420(6916):629-35 PMID: 12478284
  21. Cellular motility driven by assembly and disassembly of actin filaments.
    Cell. 2003 Feb 21;112(4):453-65 PMID: 12600310
  22. The roles of microtubule-based motor proteins in mitosis: comprehensive RNAi analysis in the Drosophila S2 cell line.
    J Cell Biol. 2003 Sep 15;162(6):1003-16 PMID: 12975346
  23. Molecular mechanism of actin-dependent retrograde flow in lamellipodia of motile cells.
    Front Biosci. 1997;2:d260-70 PMID: 9206973
  24. ASP-56, a new actin sequestering protein from pig platelets with homology to CAP, an adenylate cyclase-associated protein from yeast.
    FEBS Lett. 1992 Feb 24;298(2-3):149-53 PMID: 1544438
  25. chickadee encodes a profilin required for intercellular cytoplasm transport during Drosophila oogenesis.
    Cell. 1992 Apr 3;69(1):173-84 PMID: 1339308
  26. Improved stability of Drosophila larval neuromuscular preparations in haemolymph-like physiological solutions.
    J Comp Physiol A. 1994 Aug;175(2):179-91 PMID: 8071894
  27. An actin monomer binding activity localizes to the carboxyl-terminal half of the Saccharomyces cerevisiae cyclase-associated protein.
    J Biol Chem. 1995 Mar 10;270(10):5680-5 PMID: 7890691
  28. Profilin mutations disrupt multiple actin-dependent processes during Drosophila development.
    Development. 1994 Apr;120(4):717-28 PMID: 7600952
  29. Mutations in twinstar, a Drosophila gene encoding a cofilin/ADF homologue, result in defects in centrosome migration and cytokinesis.
    J Cell Biol. 1995 Dec;131(5):1243-59 PMID: 8522587
  30. Cell migration: a physically integrated molecular process.
    Cell. 1996 Feb 9;84(3):359-69 PMID: 8608589
  31. Actin-based cell motility and cell locomotion.
    Cell. 1996 Feb 9;84(3):371-9 PMID: 8608590
  32. Cell motility driven by actin polymerization.
    Biophys J. 1996 Dec;71(6):3030-45 PMID: 8968574
  33. ACTIN: general principles from studies in yeast.
    Annu Rev Cell Dev Biol. 1996;12:129-60 PMID: 8970724
  34. 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
  35. Scar, a WASp-related protein, activates nucleation of actin filaments by the Arp2/3 complex.
    Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3739-44 PMID: 10097107
  36. XAIP1: a Xenopus homologue of yeast actin interacting protein 1 (AIP1), which induces disassembly of actin filaments cooperatively with ADF/cofilin family proteins.
    J Cell Sci. 1999 May;112 ( Pt 10):1553-65 PMID: 10212149
  37. Aip1p interacts with cofilin to disassemble actin filaments.
    J Cell Biol. 1999 Jun 14;145(6):1251-64 PMID: 10366597
  38. The genetics of cell migration in Drosophila melanogaster and Caenorhabditis elegans development.
    Development. 1999 Jun;126(14):3035-46 PMID: 10375496
  39. Ca2+-dependent myosin II activation is required for uropod retraction during neutrophil migration.
    J Cell Sci. 2000 Apr;113 ( Pt 7):1287-98 PMID: 10704379
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200303023
Published
2003-09-15
Pages
1079-88
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2172842
Subset
IM
Grants
NIGMS NIH HHS · F32 GM064966 · United States
NIGMS NIH HHS · R01 GM038499 · United States
NIGMS NIH HHS · 5F32GM064966-02 · United States
External Links
PubMed source DOI: 10.1083/jcb.200303023 PMC Full Text View journal details
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