Ena/VASP Proteins Have an Anti-Capping Independent Function in Filopodia Formation
10.1091/mbc.e06-11-0990
Crossref journal-article
American Society for Cell Biology (ASCB)
Molecular Biology of the Cell (1076)
Abstract

Filopodia have been implicated in a number of diverse cellular processes including growth-cone path finding, wound healing, and metastasis. The Ena/VASP family of proteins has emerged as key to filopodia formation but the exact mechanism for how they function has yet to be fully elucidated. Using cell spreading as a model system in combination with small interfering RNA depletion of Capping Protein, we determined that Ena/VASP proteins have a role beyond anticapping activity in filopodia formation. Analysis of mutant Ena/VASP proteins demonstrated that the entire EVH2 domain was the minimal domain required for filopodia formation. Fluorescent recovery after photobleaching data indicate that Ena/VASP proteins rapidly exchange at the leading edge of lamellipodia, whereas virtually no exchange occurred at filopodial tips. Mutation of the G-actin–binding motif (GAB) partially compromised stabilization of Ena/VASP at filopodia tips. These observations led us to propose a model where the EVH2 domain of Ena/VASP induces and maintains clustering of the barbed ends of actin filaments, which putatively corresponds to a transition from lamellipodial to filopodial localization. Furthermore, the EVH1 domain, together with the GAB motif in the EVH2 domain, helps to maintain Ena/VASP at the growing barbed ends.

Bibliography

Applewhite, D. A., Barzik, M., Kojima, S., Svitkina, T. M., Gertler, F. B., & Borisy, G. G. (2007). Ena/VASP Proteins Have an Anti-Capping Independent Function in Filopodia Formation. Molecular Biology of the Cell, 18(7), 2579–2591.

Authors 6 University of Pennsylvania
  1. Derek A. Applewhite (first)
  2. Melanie Barzik (additional)
  3. Shin-ichiro Kojima (additional)
  4. Tatyana M. Svitkina (additional) University of Pennsylvania
  5. Frank B. Gertler (additional)
  6. Gary G. Borisy (additional)
References 72 Referenced 182
  1. 10.1083/jcb.150.4.807 / J. Cell Biol by Adams J. C. (2000)
  2. 10.1073/pnas.96.9.4977 / Proc. Natl. Acad. Sci. USA by Ahern-Djamali S. M. (1999)
  3. 10.1083/jcb.69.2.275 / J. Cell Biol by Albrecht-Buehler G. (1976)
  4. 10.1074/jbc.274.33.23549 / J. Biol. Chem by Bachmann C. (1999)
  5. 10.1242/jcs.111.12.1649 / J. Cell. Sci by Ballestrem C. (1998)
  6. 10.1016/S0955-0674(99)00059-9 / Curr. Opin. Cell Biol by Bartles J. R. (2000)
  7. 10.1074/jbc.M503957200 / J. Biol. Chem by Barzik M. (2005)
  8. 10.1016/S0092-8674(00)80884-3 / Cell by Bear J. E. (2000)
  9. 10.1016/S0092-8674(02)00731-6 / Cell by Bear J. E. (2002)
  10. 10.1002/1097-0169(200012)47:4<351::AID-CM8>3.0.CO;2-8 / Cell Motil. Cytoskelet by Bearer E. L. (2000)
  11. 10.1073/pnas.0602443103 / Proc. Natl. Acad. Sci. USA by Bohil A. B. (2006)
  12. 10.1016/S0960-9822(99)80315-7 / Curr. Biol by Carl U. D. (1999)
  13. 10.1002/j.1460-2075.1995.tb07117.x / EMBO J by Chakraborty T. (1995)
  14. 10.1016/j.jsb.2006.01.012 / J. Struct. Biol by Chereau D. (2006)
  15. 10.1016/j.cell.2006.12.049 / Cell by Co C. (2007)
  16. 10.1016/S0896-6273(03)00633-0 / Neuron by Dent E. W. (2003)
  17. 10.1016/S0006-3495(04)74246-0 / Biophys. J by Dubin-Thaler B. J. (2004)
  18. 10.1074/jbc.272.52.32869 / J. Biol. Chem by Ermekova K. S. (1997)
  19. 10.1242/jcs.00611 / J. Cell Sci by Evangelista M. (2003)
  20. 10.1038/10717 / Nat. Struct. Biol by Fedorov A. A. (1999)
  21. 10.1091/mbc.e02-01-0058
  22. 10.1101/gad.9.5.521 / Genes Dev by Gertler F. B. (1995)
  23. 10.1016/S0092-8674(00)81341-0 / Cell by Gertler F. B. (1996)
  24. 10.1074/jbc.M209107200 / J. Biol. Chem by Han Y. H. (2002)
  25. 10.1074/jbc.M005066200 / J. Biol. Chem by Harbeck B. (2000)
  26. 10.1016/j.biocel.2005.05.004 / Int. J. Biochem. Cell Biol by Hashimoto Y. (2005)
  27. 10.1073/pnas.0508269103 / Proc. Natl. Acad. Sci. USA by Haviv L. (2006)
  28. 10.1111/j.1432-1033.1994.00021.x / Eur. J. Biochem by Horstrup K. (1994)
  29. 10.1074/jbc.M205379200 / J. Biol. Chem by Howe A. K. (2002)
  30. 10.1016/S0014-5793(99)00546-3 / FEBS Lett by Huttelmaier S. (1999)
  31. 10.1038/35074643 / Nat. Cell Biol by Jacinto A. (2001)
  32. 10.1016/S0960-9822(00)00796-X / Curr. Biol by Jacinto A. (2000)
  33. 10.2144/04361ST02 / Biotechniques by Kojima S. (2004)
  34. 10.1016/j.ceb.2005.12.011 / Curr. Opin. Cell Biol by Kovar D. R. (2006)
  35. 10.1146/annurev.cellbio.19.050103.103356 / Annu. Rev. Cell Dev. Biol by Krause M. (2003)
  36. 10.1016/j.devcel.2004.07.024 / Dev. Cell by Krause M. (2004)
  37. 10.1073/pnas.0403069101 / Proc. Natl. Acad. Sci. USA by Kuhnel K. (2004)
  38. 10.1016/S0962-8924(03)00130-2 / Trends Cell Biol by Kwiatkowski A. V. (2003)
  39. 10.1074/jbc.M006274200 / J. Biol. Chem by Lambrechts A. (2000)
  40. 10.1016/S0896-6273(00)81092-2 / Neuron by Lanier L. M. (1999)
  41. 10.1083/jcb.144.6.1245 / J. Cell Biol by Laurent V. (1999)
  42. 10.1016/S0896-6273(04)00108-4 / Neuron by Lebrand C. (2004)
  43. 10.1016/j.yexcr.2007.01.023 / Exp. Cell Res by Loitto V. M. (2007)
  44. 10.1083/jcb.200309093 / J. Cell Biol by Loomis P. A. (2003)
  45. 10.1091/mbc.e01-10-0102
  46. 10.1016/j.cell.2004.07.019 / Cell by Mejillano M. R. (2004)
  47. 10.1016/S0092-8674(00)81281-7 / Cell by Mitchison T. J. (1996)
  48. 10.1016/S0006-3495(02)73897-6 / Biophys. J by Mogilner A. (2002)
  49. 10.1529/biophysj.104.056515 / Biophys. J by Mogilner A. (2005)
  50. 10.1093/emboj/16.17.5433 / EMBO J by Niebuhr K. (1997)
  51. 10.1016/j.cub.2005.01.011 / Curr. Biol by Pellegrin S. (2005)
  52. 10.1146/annurev.biophys.29.1.545 / Annu. Rev. Biophys. Biomol. Struct by Pollard T. D. (2000)
  53. 10.1016/S0092-8674(03)00120-X / Cell by Pollard T. D. (2003)
  54. 10.1084/jem.81.3.233 / J. Exp. Med by Porter K. R. (1945)
  55. 10.1002/j.1460-2075.1992.tb05264.x / EMBO J by Reinhard M. (1992)
  56. 10.1038/13040 / Nat. Cell Biol by Rottner K. (1999)
  57. 10.1083/jcb.200303191 / J. Cell Biol by Samarin S. (2003)
  58. 10.1083/jcb.143.7.1919 / J. Cell Biol by Schafer D. A. (1998)
  59. 10.1042/BST0331256 / Biochem. Soc. Trans by Schirenbeck A. (2005)
  60. 10.1073/pnas.0511243103 / Proc. Natl. Acad. Sci. USA by Schirenbeck A. (2006)
  61. 10.1016/S0892-0354(98)90010-7 / Electron. Microsc. Rev by Small J. V. (1988)
  62. 10.1083/jcb.145.5.1009 / J. Cell Biol by Svitkina T. M. (1999)
  63. 10.1083/jcb.200210174 / J. Cell Biol by Svitkina T. M. (2003)
  64. 10.1242/jcs.01215 / J. Cell Sci by Tilney L. G. (2004)
  65. 10.1016/S0955-0674(00)00177-0 / Curr. Opin. Cell Biol by Vasioukhin V. (2001)
  66. 10.1083/jcb.200603013 / J. Cell Biol by Vignjevic D. (2006)
  67. 10.1083/jcb.200208059 / J. Cell Biol by Vignjevic D. (2003)
  68. 10.1016/S0014-5793(02)03356-2 / FEBS Lett by Walders-Harbeck B. (2002)
  69. 10.1111/j.1432-1033.1987.tb13357.x / Eur. J. Biochem by Waldmann R. (1987)
  70. 10.1016/j.ceb.2003.10.019 / Curr. Opin. Cell Biol by Zigmond S. H. (2004)
  71. 10.1016/j.cub.2003.09.057 / Curr. Biol by Zigmond S. H. (2003)
  72. 10.1021/bi020379x / Biochemistry by Zimmermann J. (2002)
Dates
Type When
Created 18 years, 3 months ago (May 3, 2007, 12:42 p.m.)
Deposited 4 years ago (Aug. 12, 2021, 10:26 p.m.)
Indexed 5 days, 4 hours ago (Aug. 27, 2025, 11:49 a.m.)
Issued 18 years, 2 months ago (July 1, 2007)
Published 18 years, 2 months ago (July 1, 2007)
Published Print 18 years, 2 months ago (July 1, 2007)
Funders 0

None

@article{Applewhite_2007, title={Ena/VASP Proteins Have an Anti-Capping Independent Function in Filopodia Formation}, volume={18}, ISSN={1939-4586}, url={http://dx.doi.org/10.1091/mbc.e06-11-0990}, DOI={10.1091/mbc.e06-11-0990}, number={7}, journal={Molecular Biology of the Cell}, publisher={American Society for Cell Biology (ASCB)}, author={Applewhite, Derek A. and Barzik, Melanie and Kojima, Shin-ichiro and Svitkina, Tatyana M. and Gertler, Frank B. and Borisy, Gary G.}, editor={Drubin, David}, year={2007}, month=jul, pages={2579–2591} }