A functional link between dynamin and the actin cytoskeleton at podosomes

Gian Carlo Ochoa, Vladimir I. Slepnev, Lynn Neff, Niels Ringstad, Kohji Takei, Laurie Daniell, Warren Kim, Hong Cao, Mark McNiven, Roland Baron, Pietro De Camilli

Research output: Contribution to journalArticle

295 Scopus citations

Abstract

Cell transformation by Rous sarcoma virus results in a dramatic change of adhesion structures with the substratum. Adhesion plaques are replaced by dot-like attachment sites called podosomes. Podosomes are also found constitutively in motile nontransformed cells such as leukocytes, macrophages, and osteoclasts. They are represented by columnar arrays of actin which are perpendicular to the substratum and contain tubular invaginations of the plasma membrane. Given the similarity of these tubules to those generated by dynamin around a variety of membrane templates, we investigated whether dynamin is present at podosomes. Immunoreactivities for dynamin 2 and for the dynamin 2-binding protein endophilin 2 (SH3P8) were detected at podosomes of transformed cells and osteoclasts. Furthermore, GFP wild-type dynamin 2aa was targeted to podosomes. As shown by fluorescence recovery after photobleaching, GFP-dynamin 2aa and GFP-actin had a very rapid and similar turnover at podosomes. Expression of the GFP-dynamin 2aa(G273D) abolished podosomes while GFP-dynamin(K44A) was targeted to podosomes but delayed actin turnover. These data demonstrate a functional link between a member of the dynamin family and actin at attachment sites between cells and the substratum.

Original languageEnglish (US)
Pages (from-to)377-389
Number of pages13
JournalJournal of Cell Biology
Volume150
Issue number2
DOIs
StatePublished - Jul 24 2000

Keywords

  • Actin patches
  • Clathrin
  • Endocytosis
  • Osteoclast
  • Src

ASJC Scopus subject areas

  • Cell Biology

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    Ochoa, G. C., Slepnev, V. I., Neff, L., Ringstad, N., Takei, K., Daniell, L., Kim, W., Cao, H., McNiven, M., Baron, R., & De Camilli, P. (2000). A functional link between dynamin and the actin cytoskeleton at podosomes. Journal of Cell Biology, 150(2), 377-389. https://doi.org/10.1083/jcb.150.2.377