Structural and functional dissection of the Abp1 ADFH actin-binding domain reveals versatile in vivo adapter functions

Omar Quintero-Monzon, Avital A. Rodal, Boris Strokopytov, Steven C. Almo, Bruce L. Goode

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Abp1 is a multidomain protein that regulates the Arp2/3 complex and links proteins involved in endocytosis to the actin cytoskeleton. All of the proposed cellular functions of Abp1 involve actin filament binding, yet the actin binding site(s) on Abp1 have not been identified, nor has the importance of actin binding for Abp1 localization and function in vivo been tested. Here, we report the crystal structure of the Saccharomyces cerevisiae Abp1 actin-binding actin depolymerizing factor homology (ADFH) domain and dissect its activities by mutagenesis. Abp1-ADFH domain and ADF/cofilin structures are similar, and they use conserved surfaces to bind actin; however, there are also key differences that help explain their differential effects on actin dynamics. Using point mutations, we demonstrate that actin binding is required for localization of Abp1 in vivo, the lethality caused by Abp1 overexpression, and the ability of Abp1 to activate Arp2/3 complex. Furthermore, we genetically uncouple ABP1 functions that overlap with SAC6, SLA1, and SLA2, showing they require distinct combinations of activities and interactions. Together, our data provide the first structural and functional view of the Abp1-actin interaction and show that Abp1 has distinct cellular roles as an adapter, linking different sets of ligands for each function.

Original languageEnglish (US)
Pages (from-to)3128-3139
Number of pages12
JournalMolecular Biology of the Cell
Volume16
Issue number7
DOIs
StatePublished - Jul 2005

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Destrin
Dissection
Actins
Actin-Related Protein 2-3 Complex
Actin Cytoskeleton
Actin Depolymerizing Factors
Endocytosis
Point Mutation
Mutagenesis
Saccharomyces cerevisiae
Binding Sites
Ligands

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Structural and functional dissection of the Abp1 ADFH actin-binding domain reveals versatile in vivo adapter functions. / Quintero-Monzon, Omar; Rodal, Avital A.; Strokopytov, Boris; Almo, Steven C.; Goode, Bruce L.

In: Molecular Biology of the Cell, Vol. 16, No. 7, 07.2005, p. 3128-3139.

Research output: Contribution to journalArticle

Quintero-Monzon, Omar ; Rodal, Avital A. ; Strokopytov, Boris ; Almo, Steven C. ; Goode, Bruce L. / Structural and functional dissection of the Abp1 ADFH actin-binding domain reveals versatile in vivo adapter functions. In: Molecular Biology of the Cell. 2005 ; Vol. 16, No. 7. pp. 3128-3139.
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