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 A Mc Niven, Roland Baron, Pietro De Camilli

Research output: Contribution to journalArticle

292 Citations (Scopus)

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

Fingerprint

Dynamins
Actin Cytoskeleton
Actins
Dynamin II
Osteoclasts
Fluorescence Recovery After Photobleaching
Rous sarcoma virus
Focal Adhesions
Podosomes
Carrier Proteins
Leukocytes
Macrophages
Cell Membrane
Membranes

Keywords

  • Actin patches
  • Clathrin
  • Endocytosis
  • Osteoclast
  • Src

ASJC Scopus subject areas

  • Cell Biology

Cite this

Ochoa, G. C., Slepnev, V. I., Neff, L., Ringstad, N., Takei, K., Daniell, L., ... 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

A functional link between dynamin and the actin cytoskeleton at podosomes. / Ochoa, Gian Carlo; Slepnev, Vladimir I.; Neff, Lynn; Ringstad, Niels; Takei, Kohji; Daniell, Laurie; Kim, Warren; Cao, Hong; Mc Niven, Mark A; Baron, Roland; De Camilli, Pietro.

In: Journal of Cell Biology, Vol. 150, No. 2, 24.07.2000, p. 377-389.

Research output: Contribution to journalArticle

Ochoa, GC, Slepnev, VI, Neff, L, Ringstad, N, Takei, K, Daniell, L, Kim, W, Cao, H, Mc Niven, MA, Baron, R & De Camilli, P 2000, 'A functional link between dynamin and the actin cytoskeleton at podosomes', Journal of Cell Biology, vol. 150, no. 2, pp. 377-389. https://doi.org/10.1083/jcb.150.2.377
Ochoa GC, Slepnev VI, Neff L, Ringstad N, Takei K, Daniell L et al. A functional link between dynamin and the actin cytoskeleton at podosomes. Journal of Cell Biology. 2000 Jul 24;150(2):377-389. https://doi.org/10.1083/jcb.150.2.377
Ochoa, Gian Carlo ; Slepnev, Vladimir I. ; Neff, Lynn ; Ringstad, Niels ; Takei, Kohji ; Daniell, Laurie ; Kim, Warren ; Cao, Hong ; Mc Niven, Mark A ; Baron, Roland ; De Camilli, Pietro. / A functional link between dynamin and the actin cytoskeleton at podosomes. In: Journal of Cell Biology. 2000 ; Vol. 150, No. 2. pp. 377-389.
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