Rabphilin localizes with the cell actin cytoskeleton and stimulates association of granules with F-actin cross-linked by α-actinin

Giovanna Baldini, Alberto M. Martelli, Giovanna Tabellini, Chad Horn, Khaled Machaca, Paola Narducci, Giulia Baldini

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

In endocrine cell, granules accumulate within an F-actin-rich region below the plasma membrane. The mechanisms involved in this process are largely unknown. Rabphilin is a cytosolic protein that is expressed in neurons and neuroendocrine cells and binds with high affinity to members of the Rab3 family of GTPases localized to synaptic vesicles and dense core granules. Rabphilin also interacts with α-actinin, a protein that cross-links F-actin into bundles and networks and associates with the granule membrane. Here we asked whether rabphilin, in addition to its granule localization, also interacts with the cell actin cytoskeleton. Immunofluorescence and immunoelectron microscopy show that rabphilin localizes to the sub-plasmalemmal actin cytoskeleton both in neuroendocrine and unspecialized cells. By using purified components, it is found that association of rabphilin with F-actin is dependent on added α-actinin. In an in vitro assay, granules, unlike endosomes or mitochondria, associate with F-actin cross-linked by α-actinin. Rabphilin is shown to stimulate this process. Rabphilin enhances by ∼8-fold the granule ability to localize within regions of elevated concentration of cross-linked F-actin. These results suggest that rabphilin, by interacting with á-actinin, organizes the cell cytoskeleton to facilitate granule localization within F-actin-rich regions.

Original languageEnglish
Pages (from-to)34974-34984
Number of pages11
JournalJournal of Biological Chemistry
Volume280
Issue number41
DOIs
Publication statusPublished - 14 Oct 2005
Externally publishedYes

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Actinin
Actin Cytoskeleton
Actins
Association reactions
Neuroendocrine Cells
Aptitude
Endocrine Cells
Immunoelectron Microscopy
Synaptic Vesicles
Endosomes
GTP Phosphohydrolases
Cytoskeleton
Fluorescence Microscopy
Mitochondria
Cell membranes
Cell Membrane
Neurons
Assays
Microscopic examination
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Rabphilin localizes with the cell actin cytoskeleton and stimulates association of granules with F-actin cross-linked by α-actinin. / Baldini, Giovanna; Martelli, Alberto M.; Tabellini, Giovanna; Horn, Chad; Machaca, Khaled; Narducci, Paola; Baldini, Giulia.

In: Journal of Biological Chemistry, Vol. 280, No. 41, 14.10.2005, p. 34974-34984.

Research output: Contribution to journalArticle

Baldini, Giovanna ; Martelli, Alberto M. ; Tabellini, Giovanna ; Horn, Chad ; Machaca, Khaled ; Narducci, Paola ; Baldini, Giulia. / Rabphilin localizes with the cell actin cytoskeleton and stimulates association of granules with F-actin cross-linked by α-actinin. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 41. pp. 34974-34984.
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