title: Eng2 is a component of a dynamic protein complex required for endocytic uptake in fission yeast.
abstract: Eng2 is a glucanase required for spore release, although it is also expressed during vegetative growth, suggesting that it might play other cellular functions. Its homology to the Saccharomyces cerevisiae Acf2 protein, previously shown to promote actin polymerization at endocytic sites in vitro, prompted us to investigate its role in endocytosis. Interestingly, depletion of Eng2 caused profound defects in endocytic uptake, which were not due to the absence of its glucanase activity. Analysis of the dynamics of endocytic proteins by fluorescence microscopy in the eng2Δ strain unveiled a previously undescribed phenotype, in which assembly of the Arp2/3 complex appeared uncoupled from the internalization of the endocytic coat and resulted in a fission defect. Strikingly also, we found that Eng2-GFP dynamics did not match the pattern of other endocytic proteins. Eng2-GFP localized to bright cytosolic spots that moved around the cellular poles and occasionally contacted assembling endocytic patches just before recruitment of Wsp1, the Schizosaccharomyces pombe WASP. Interestingly, Csh3-YFP, a WASP-interacting protein, interacted with Eng2 by co-immunoprecipitation and was recruited to Eng2 in bright cytosolic spots. Altogether, our work defines a novel endocytic functional module, which probably couples the endocytic coat to the actin module.
authors.affiliation: Instituto de Biología Funcional y Genómica, CSIC/Universidad de Salamanca, c/ Zacarías González 2, 37007, Salamanca, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Encinar del Dedo
authors.forename: Fatima-Zahra
authors.initials: FZ
authors.lastname: Idrissi
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Arnáiz-Pita
authors.forename: Michael
authors.initials: M
authors.lastname: James
authors.forename: Encarnación
authors.initials: E
authors.lastname: Dueñas-Santero
authors.forename: Sara
authors.initials: S
authors.lastname: Orellana-Muñoz
authors.forename: Francisco
authors.initials: F
authors.lastname: del Rey
authors.forename: Vladimir
authors.initials: V
authors.lastname: Sirotkin
authors.forename: M Isabel
authors.initials: MI
authors.lastname: Geli
authors.forename: Carlos R
authors.initials: CR
authors.lastname: Vázquez de Aldana
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D051376:Actin-Related Protein 2-3 Complex / Q000378:metabolism; D004705:Endocytosis; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D051376:Actin-Related Protein 2-3 Complex; D029702:Schizosaccharomyces pombe Proteins; C417034:Wsp1 protein, S pombe; C000599813:Eng2 protein, S pombe; D004693:Glucan Endo-1,3-beta-D-Glucosidase
keywords: Arp2-3 complex; End4-Sla2; Wsp1; actin polymerization; endocytosis