title: A nanobody-based toolset to investigate the role of protein localization and dispersal inDrosophila.
abstract: The role of protein localization along the apical-basal axis of polarized cells is difficult to investigate in vivo, partially due to lack of suitable tools. Here, we present the GrabFP system, a collection of four nanobody-based GFP-traps that localize to defined positions along the apical-basal axis. We show that the localization preference of the GrabFP traps can impose a novel localization on GFP-tagged target proteins and results in their controlled mislocalization. These new tools were used to mislocalize transmembrane and cytoplasmic GFP fusion proteins in theDrosophilawing disc epithelium and to investigate the effect of protein mislocalization. Furthermore, we used the GrabFP system as a tool to study the extracellular dispersal of the Decapentaplegic (Dpp) protein and show that the Dpp gradient forming in the lateral plane of theDrosophilawing disc epithelium is essential for patterning of the wing imaginal disc.
authors.affiliation: Growth and Development, Biozentrum, University of Basel, Basel, Switzerland.
authors.forename: Stefan
authors.identifier: 0000-0001-6668-7608
authors.initials: S
authors.lastname: Harmansa
authors.affiliation: Growth and Development, Biozentrum, University of Basel, Basel, Switzerland.
authors.forename: Ilaria
authors.identifier: 0000-0001-7849-3958
authors.initials: I
authors.lastname: Alborelli
authors.affiliation: Growth and Development, Biozentrum, University of Basel, Basel, Switzerland.
authors.forename: Dimitri
authors.initials: D
authors.lastname: Bieli
authors.affiliation: Growth and Development, Biozentrum, University of Basel, Basel, Switzerland.|Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Caussinus
authors.affiliation: Growth and Development, Biozentrum, University of Basel, Basel, Switzerland.
authors.forename: Markus
authors.identifier: 0000-0002-5171-0016
authors.initials: M
authors.lastname: Affolter
mesh_terms: D000818:Animals; D004330:Drosophila / Q000196:embryology* / Q000235:genetics / Q000502:physiology; D029721:Drosophila Proteins / Q000378:metabolism*; D004772:Entomology / Q000379:methods*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D061905:Single-Domain Antibodies / Q000378:metabolism*
chemical_list: D029721:Drosophila Proteins; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies; D049452:Green Fluorescent Proteins
keywords: D. melanogaster; Dpp; cell biology; developmental biology; morphogen; nanobody; protein localization; stem cells