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PMID: 22232703 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

α-Actinin-4/FSGS1 is required for Arp2/3-dependent actin assembly at the adherens junction.

The Journal of cell biology ·Vol. 196 ·No. 1 ·2012-01-09 ·Pages 115-30

Tang VW, Brieher WM

Abstract

We have developed an in vitro assay to study actin assembly at cadherin-enriched cell junctions. Using this assay, we demonstrate that cadherin-enriched junctions can polymerize new actin filaments but cannot capture preexisting filaments, suggesting a mechanism involving de novo synthesis. In agreement with this hypothesis, inhibition of Arp2/3-dependent nucleation abolished actin assembly at cell-cell junctions. Reconstitution biochemistry using the in vitro actin assembly assay identified α-actinin-4/focal segmental glomerulosclerosis 1 (FSGS1) as an essential factor. α-Actinin-4 specifically localized to sites of actin incorporation on purified membranes and at apical junctions in Madin-Darby canine kidney cells. Knockdown of α-actinin-4 decreased total junctional actin and inhibited actin assembly at the apical junction. Furthermore, a point mutation of α-actinin-4 (K255E) associated with FSGS failed to support actin assembly and acted as a dominant negative to disrupt actin dynamics at junctional complexes. These findings demonstrate that α-actinin-4 plays an important role in coupling actin nucleation to assembly at cadherin-based cell-cell adhesive contacts.

MeSH Terms
Actin Cytoskeleton/metabolism Actin-Related Protein 2-3 Complex/physiology Actinin/genetics,physiology Actins/analysis Adherens Junctions/metabolism Animals Cadherins/metabolism Cell Adhesion/genetics Cell Line Dogs Point Mutation Polymerization
Chemicals
Actin-Related Protein 2-3 Complex Actins Cadherins Actinin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Tang Vivian W
Department of Cell and Developmental Biology, University of Illinois, Urbana, IL 61801, USA.
Brieher William M
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.201103116
Published
2012-01-09
Pages
115-30
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC3255975
Subset
IM
Grants
NIDDK NIH HHS · R01 DK098398 · United States
External Links
PubMed source DOI: 10.1083/jcb.201103116 PMC Full Text View journal details
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