title: Drosophila homologues of adenomatous polyposis coli (APC) and the formin diaphanous collaborate by a conserved mechanism to stimulate actin filament assembly.
abstract: BACKGROUND
  Vertebrate APC collaborates with Dia through its Basic domain to assemble actin filaments.
  
  
  RESULTS
  Despite limited sequence homology between the vertebrate and Drosophila APC Basic domains, Drosophila APC1 collaborates with Dia to stimulate actin assembly in vitro.
  
  
  CONCLUSIONS
  The mechanism of actin assembly is highly conserved over evolution.
  
  
  CONCLUSIONS
  APC-Dia collaborations may be crucial in a wide range of animal cells. Adenomatous polyposis coli (APC) is a large multidomain protein that regulates the cytoskeleton. Recently, it was shown that vertebrate APC through its Basic domain directly collaborates with the formin mDia1 to stimulate actin filament assembly in the presence of nucleation barriers. However, it has been unclear whether these activities extend to homologues of APC and Dia in other organisms. Drosophila APC and Dia are each required to promote actin furrow formation in the syncytial embryo, suggesting a potential collaboration in actin assembly, but low sequence homology between the Basic domains of Drosophila and vertebrate APC has left their functional and mechanistic parallels uncertain. To address this question, we purified Drosophila APC1 and Dia and determined their individual and combined effects on actin assembly using both bulk fluorescence assays and total internal reflection fluorescence microscopy. Our data show that APC1, similar to its vertebrate homologue, bound to actin monomers and nucleated and bundled filaments. Further, Drosophila Dia nucleated actin assembly and protected growing filament barbed ends from capping protein. Drosophila APC1 and Dia directly interacted and collaborated to promote actin assembly in the combined presence of profilin and capping protein. Thus, despite limited sequence homology, Drosophila and vertebrate APCs exhibit highly related activities and mechanisms and directly collaborate with formins. These results suggest that APC-Dia interactions in actin assembly are conserved and may underlie important in vivo functions in a broad range of animal phyla.
authors.affiliation: Department of Biology, Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA 02454, USA.
authors.forename: Richa
authors.initials: R
authors.lastname: Jaiswal
authors.forename: Vince
authors.initials: V
authors.lastname: Stepanik
authors.forename: Aneliya
authors.initials: A
authors.lastname: Rankova
authors.forename: Olivia
authors.initials: O
authors.lastname: Molinar
authors.forename: Bruce L
authors.initials: BL
authors.lastname: Goode
authors.forename: Brooke M
authors.initials: BM
authors.lastname: McCartney
mesh_terms: D000199:Actins / Q000737:chemistry*; D000818:Animals; D051347:CapZ Actin Capping Protein / Q000737:chemistry; D002352:Carrier Proteins / Q000737:chemistry*; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins / Q000737:chemistry*; D004331:Drosophila melanogaster; D000080865:Formins; D007700:Kinetics; D010446:Peptide Fragments / Q000737:chemistry; D051304:Profilins / Q000737:chemistry; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D025521:Tumor Suppressor Proteins / Q000737:chemistry*
chemical_list: C118001:APC protein, Drosophila; D000199:Actins; D051347:CapZ Actin Capping Protein; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins; D000080865:Formins; D010446:Peptide Fragments; D051304:Profilins; D025521:Tumor Suppressor Proteins; C106890:diaphanous protein, Drosophila
keywords: APC1B; Actin; Drosophila; Fluorescence; Formin; Protein Purification; Protein-Protein Interactions; TIRF Microscopy
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