A barbed end interference mechanism reveals how capping protein promotes nucleation in branched actin networks.
Actin Capping Proteins
/ chemistry
Actin Cytoskeleton
/ metabolism
Actin-Related Protein 2-3 Complex
/ chemistry
Actins
/ chemistry
Animals
Binding Sites
Cattle
Cytoskeleton
/ metabolism
Gelsolin
/ chemistry
Gene Expression Regulation
Humans
Intercellular Signaling Peptides and Proteins
/ chemistry
Kinetics
Melanocytes
/ cytology
Melanoma, Experimental
/ genetics
Mice
Models, Molecular
Profilins
/ chemistry
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Recombinant Proteins
/ chemistry
Thymus Gland
/ cytology
Wiskott-Aldrich Syndrome Protein, Neuronal
/ chemistry
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
09 09 2021
09 09 2021
Historique:
received:
10
04
2021
accepted:
18
08
2021
entrez:
10
9
2021
pubmed:
11
9
2021
medline:
7
10
2021
Statut:
epublish
Résumé
Heterodimeric capping protein (CP/CapZ) is an essential factor for the assembly of branched actin networks, which push against cellular membranes to drive a large variety of cellular processes. Aside from terminating filament growth, CP potentiates the nucleation of actin filaments by the Arp2/3 complex in branched actin networks through an unclear mechanism. Here, we combine structural biology with in vitro reconstitution to demonstrate that CP not only terminates filament elongation, but indirectly stimulates the activity of Arp2/3 activating nucleation promoting factors (NPFs) by preventing their association to filament barbed ends. Key to this function is one of CP's C-terminal "tentacle" extensions, which sterically masks the main interaction site of the terminal actin protomer. Deletion of the β tentacle only modestly impairs capping. However, in the context of a growing branched actin network, its removal potently inhibits nucleation promoting factors by tethering them to capped filament ends. End tethering of NPFs prevents their loading with actin monomers required for activation of the Arp2/3 complex and thus strongly inhibits branched network assembly both in cells and reconstituted motility assays. Our results mechanistically explain how CP couples two opposed processes-capping and nucleation-in branched actin network assembly.
Identifiants
pubmed: 34504078
doi: 10.1038/s41467-021-25682-5
pii: 10.1038/s41467-021-25682-5
pmc: PMC8429771
doi:
Substances chimiques
Actin Capping Proteins
0
Actin-Related Protein 2-3 Complex
0
Actins
0
Gelsolin
0
Intercellular Signaling Peptides and Proteins
0
Profilins
0
Recombinant Proteins
0
WASL protein, human
0
Wiskott-Aldrich Syndrome Protein, Neuronal
0
myotrophin
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
5329Informations de copyright
© 2021. The Author(s).
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