Characterization of the biochemical properties and biological function of the formin homology domains of Drosophila DAAM

Szilvia Barko, B. Bugyi, Marie France Carlier, Rita Gombos, Tamás Matusek, József Mihályand, M. Nyitrai

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Abstract

We characterized the properties of Drosophila melanogaster DAAM-FH2 and DAAM-FH1-FH2 fragments and their interactions with actin and profilin by using various biophysical methods and in vivo experiments. The results show that although the DAAM-FH2 fragment does not have any conspicuous effect on actin assembly in vivo, in cells expressing the DAAM-FH1-FH2 fragment, a profilin-dependent increase in the formation of actin structures is observed. The trachea-specific expression of DAAM-FH1-FH2also induces phenotypic effects, leading to the collapse of the tracheal tube and lethality in the larval stages. In vitro, both DAAM fragments catalyze actin nucleation but severely decrease both the elongation and depolymerization rate of the filaments. Profilin acts as a molecular switch in DAAMfunction. DAAM-FH1-FH2, remaining bound to barbed ends, drives processive assembly of profilin-actin, whereas DAAM-FH2 forms an abortive complex with barbed ends that does not support profilin-actin assembly. Both DAAM fragments also bind to the sides of the actin filaments and induce actin bundling. These observations show that the D. melanogaster DAAM formin represents an extreme class of barbed end regulators gated by profilin.

Original languageEnglish
Pages (from-to)13154-13169
Number of pages16
JournalJournal of Biological Chemistry
Volume285
Issue number17
DOIs
Publication statusPublished - Apr 23 2010

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Profilins
Drosophila
Actins
Drosophila melanogaster
Depolymerization
Trachea
Actin Cytoskeleton
Elongation
Nucleation
Cells
Switches

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Characterization of the biochemical properties and biological function of the formin homology domains of Drosophila DAAM. / Barko, Szilvia; Bugyi, B.; Carlier, Marie France; Gombos, Rita; Matusek, Tamás; Mihályand, József; Nyitrai, M.

In: Journal of Biological Chemistry, Vol. 285, No. 17, 23.04.2010, p. 13154-13169.

Research output: Contribution to journalArticle

Barko, Szilvia ; Bugyi, B. ; Carlier, Marie France ; Gombos, Rita ; Matusek, Tamás ; Mihályand, József ; Nyitrai, M. / Characterization of the biochemical properties and biological function of the formin homology domains of Drosophila DAAM. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 17. pp. 13154-13169.
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