title: Structural requirements for PACSIN/Syndapin operation during zebrafish embryonic notochord development.
abstract: PACSIN/Syndapin proteins are membrane-active scaffolds that participate in endocytosis. The structure of the Drosophila Syndapin N-terminal EFC domain reveals a crescent shaped antiparallel dimer with a high affinity for phosphoinositides and a unique membrane-inserting prong upon the concave surface. Combined structural, biochemical and reverse genetic approaches in zebrafish define an important role for Syndapin orthologue, Pacsin3, in the early formation of the notochord during embryonic development. In pacsin3-morphant embryos, midline convergence of notochord precursors is defective as axial mesodermal cells fail to polarize, migrate and differentiate properly. The pacsin3 morphant phenotype of a stunted body axis and contorted trunk is rescued by ectopic expression of Drosophila Syndapin, and depends critically on both the prong that protrudes from the surface of the bowed Syndapin EFC domain and the ability of the antiparallel dimer to bind tightly to phosphoinositides. Our data confirm linkage between directional migration, endocytosis and cell specification during embryonic morphogenesis and highlight a key role for Pacsin3 in this coupling in the notochord.
authors.affiliation: Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Edeling
authors.forename: Subramaniam
authors.initials: S
authors.lastname: Sanker
authors.forename: Takaki
authors.initials: T
authors.lastname: Shima
authors.forename: P K
authors.initials: PK
authors.lastname: Umasankar
authors.forename: Stefan
authors.initials: S
authors.lastname: Höning
authors.forename: Hye Y
authors.initials: HY
authors.lastname: Kim
authors.forename: Lance A
authors.initials: LA
authors.lastname: Davidson
authors.forename: Simon C
authors.initials: SC
authors.lastname: Watkins
authors.forename: Michael
authors.initials: M
authors.lastname: Tsang
authors.forename: David J
authors.initials: DJ
authors.lastname: Owen
authors.forename: Linton M
authors.initials: LM
authors.lastname: Traub
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D002465:Cell Movement / Q000187:drug effects; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000473:pathology / Q000648:ultrastructure; D047108:Embryonic Development / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D008969:Molecular Sequence Data; D009672:Notochord / Q000002:abnormalities / Q000187:drug effects / Q000196:embryology* / Q000473:pathology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010641:Phenotype; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D015027:Zebrafish / Q000196:embryology*; D029961:Zebrafish Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D029961:Zebrafish Proteins; C547742:pacsin3 protein, zebrafish