2.50
Hdl Handle:
http://hdl.handle.net/10033/120629
Title:
Molecular basis for the dual function of Eps8 on actin dynamics: bundling and capping.
Authors:
Hertzog, Maud; Milanesi, Francesca; Hazelwood, Larnele; Disanza, Andrea; Liu, HongJun; Perlade, Emilie; Malabarba, Maria Grazia; Pasqualato, Sebastiano; Maiolica, Alessio; Confalonieri, Stefano; Le Clainche, Christophe; Offenhauser, Nina; Block, Jennifer; Rottner, Klemens; Di Fiore, Pier Paolo; Carlier, Marie-France; Volkmann, Niels; Hanein, Dorit; Scita, Giorgio
Abstract:
Actin capping and cross-linking proteins regulate the dynamics and architectures of different cellular protrusions. Eps8 is the founding member of a unique family of capping proteins capable of side-binding and bundling actin filaments. However, the structural basis through which Eps8 exerts these functions remains elusive. Here, we combined biochemical, molecular, and genetic approaches with electron microscopy and image analysis to dissect the molecular mechanism responsible for the distinct activities of Eps8. We propose that bundling activity of Eps8 is mainly mediated by a compact four helix bundle, which is contacting three actin subunits along the filament. The capping activity is mainly mediated by a amphipathic helix that binds within the hydrophobic pocket at the barbed ends of actin blocking further addition of actin monomers. Single-point mutagenesis validated these modes of binding, permitting us to dissect Eps8 capping from bundling activity in vitro. We further showed that the capping and bundling activities of Eps8 can be fully dissected in vivo, demonstrating the physiological relevance of the identified Eps8 structural/functional modules. Eps8 controls actin-based motility through its capping activity, while, as a bundler, is essential for proper intestinal morphogenesis of developing Caenorhabditis elegans.
Affiliation:
IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Milan, Italy.
Citation:
Molecular basis for the dual function of Eps8 on actin dynamics: bundling and capping. 2010, 8 (6):e1000387 PLoS Biol.
Journal:
PLoS biology
Issue Date:
2010
URI:
http://hdl.handle.net/10033/120629
DOI:
10.1371/journal.pbio.1000387
PubMed ID:
20532239
Type:
Article
Language:
en
ISSN:
1545-7885
Appears in Collections:
Publications of RG Cytoskeleton Dynamics (CYD)

Full metadata record

DC FieldValue Language
dc.contributor.authorHertzog, Mauden
dc.contributor.authorMilanesi, Francescaen
dc.contributor.authorHazelwood, Larneleen
dc.contributor.authorDisanza, Andreaen
dc.contributor.authorLiu, HongJunen
dc.contributor.authorPerlade, Emilieen
dc.contributor.authorMalabarba, Maria Graziaen
dc.contributor.authorPasqualato, Sebastianoen
dc.contributor.authorMaiolica, Alessioen
dc.contributor.authorConfalonieri, Stefanoen
dc.contributor.authorLe Clainche, Christopheen
dc.contributor.authorOffenhauser, Ninaen
dc.contributor.authorBlock, Jenniferen
dc.contributor.authorRottner, Klemensen
dc.contributor.authorDi Fiore, Pier Paoloen
dc.contributor.authorCarlier, Marie-Franceen
dc.contributor.authorVolkmann, Nielsen
dc.contributor.authorHanein, Doriten
dc.contributor.authorScita, Giorgioen
dc.date.accessioned2011-01-28T15:52:02Z-
dc.date.available2011-01-28T15:52:02Z-
dc.date.issued2010-
dc.identifier.citationMolecular basis for the dual function of Eps8 on actin dynamics: bundling and capping. 2010, 8 (6):e1000387 PLoS Biol.en
dc.identifier.issn1545-7885-
dc.identifier.pmid20532239-
dc.identifier.doi10.1371/journal.pbio.1000387-
dc.identifier.urihttp://hdl.handle.net/10033/120629-
dc.description.abstractActin capping and cross-linking proteins regulate the dynamics and architectures of different cellular protrusions. Eps8 is the founding member of a unique family of capping proteins capable of side-binding and bundling actin filaments. However, the structural basis through which Eps8 exerts these functions remains elusive. Here, we combined biochemical, molecular, and genetic approaches with electron microscopy and image analysis to dissect the molecular mechanism responsible for the distinct activities of Eps8. We propose that bundling activity of Eps8 is mainly mediated by a compact four helix bundle, which is contacting three actin subunits along the filament. The capping activity is mainly mediated by a amphipathic helix that binds within the hydrophobic pocket at the barbed ends of actin blocking further addition of actin monomers. Single-point mutagenesis validated these modes of binding, permitting us to dissect Eps8 capping from bundling activity in vitro. We further showed that the capping and bundling activities of Eps8 can be fully dissected in vivo, demonstrating the physiological relevance of the identified Eps8 structural/functional modules. Eps8 controls actin-based motility through its capping activity, while, as a bundler, is essential for proper intestinal morphogenesis of developing Caenorhabditis elegans.en
dc.language.isoenen
dc.subject.meshActinsen
dc.subject.meshHumansen
dc.subject.meshIntracellular Signaling Peptides and Proteinsen
dc.subject.meshMass Spectrometryen
dc.subject.meshMicroscopy, Electronen
dc.subject.meshModels, Molecularen
dc.subject.meshProtein Bindingen
dc.subject.meshThermodynamicsen
dc.titleMolecular basis for the dual function of Eps8 on actin dynamics: bundling and capping.en
dc.typeArticleen
dc.contributor.departmentIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Milan, Italy.en
dc.identifier.journalPLoS biologyen

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