title: The function of a Drosophila glypican does not depend entirely on heparan sulfate modification.
abstract: Division abnormally delayed (Dally) is one of two glycosylphosphatidylinositol (GPI)-linked heparan sulfate proteoglycans in Drosophila. Numerous studies have shown that it influences Decapentaplegic (Dpp) and Wingless signaling. It has been generally assumed that Dally affects signaling by directly interacting with these growth factors, primarily through its heparan sulfate (HS) chains. To understand the functional contributions of HS chains and protein core we have (1) assessed the growth factor binding properties of purified Dally using surface plasmon resonance, (2) generated a form of Dally that is not HS modified and evaluated its signaling capacity in vivo. Purified Dally binds directly to FGF2, FGF10, and the functional Dpp homolog BMP4. FGF binding is abolished by preincubation with HS, but BMP4 association is partially HS-resistant, suggesting the Dally protein core contributes to binding. Cell binding and co-immunoprecipitation studies suggest that non-HS-modified Dally retains some ability to bind Dpp or BMP4. Expression of HS-deficient Dally in vivo showed it does not promote signaling as well as wild-type Dally, yet it can rescue several dally mutant phenotypes. These data reveal that heparan sulfate modification of Dally is not required for all in vivo activities and that significant functional capacity resides in the protein core.
authors.affiliation: The Developmental Biology Center, Department of Pediatrics, 6-160 Jackson Hall, 321 Church Street SE, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Kirkpatrick
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Knox
authors.forename: William D
authors.initials: WD
authors.lastname: Staatz
authors.forename: Bethany
authors.initials: B
authors.lastname: Fox
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Lercher
authors.forename: Scott B
authors.initials: SB
authors.lastname: Selleck
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D029721:Drosophila Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D053673:Glypicans / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D006497:Heparan Sulfate / Q000737:chemistry / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011509:Proteoglycans / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D013329:Structure-Activity Relationship
chemical_list: D029721:Drosophila Proteins; D053673:Glypicans; D006497:Heparan Sulfate; D008562:Membrane Glycoproteins; D011509:Proteoglycans; C098128:dally protein, Drosophila