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Drosophila GPI-mannosyltransferase 2 is required for GPI anchor attachment and surface expression of chaoptin

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  • Published online: 10 May 2012

Glycosylphosphatidylinositol (GPI) anchors are critical for the membrane attachment of a wide variety of essential signaling and cell adhesion proteins. The GPI anchor is a complex glycolipid structure that utilizes glycosylphosphatidylinositol-mannosyltransferases (GPI-MTs) for the addition of three core mannose residues during its biosynthesis. Here, we demonstrate that Drosophila GPI-MT2 is required for the GPI-mediated membrane attachment of several GPI-anchored proteins, including the photoreceptor-specific cell adhesion molecule, chaoptin. Mutations in gpi-mt2 lead to defects in chaoptin trafficking to the plasma membrane in Drosophila photoreceptor cells. In gpi-mt2 mutants, loss of sufficient chaoptin in the membrane leads to microvillar instability, photoreceptor cell pathology, and retinal degeneration. Finally, using site-directed mutagenesis, we have identified key amino acids that are essential for GPI-MT2 function and cell viability in Drosophila. Our findings on GPI-MT2 provide a mechanistic link between GPI anchor biosynthesis and protein trafficking in Drosophila and shed light on a novel mechanism for inherited retinal degeneration.

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*Address correspondence and reprint requests to: Dr. Nansi Jo Colley, Department of Ophthalmology & Visual Sciences, K6/460, 600 Highland Avenue, Madison, WI, 53792. E-mail:
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A.M. Almeida , Y. Murakami , D.M. Layton , P. Hillmen , G.S. Sellick , Y. Maeda , S. Richards , S. Patterson , I. Kotsianidis , L. Mollica , D.H. Crawford , A. Baker , M. Ferguson , I. Roberts , R. Houlston , T. Kinoshita & A. Karadimitris (2006). Hypomorphic promoter mutation in PIGM causes inherited glycosylphosphatidylinositol deficiency. Nature Medicine 12, 846851.

R.B. Arrighi & I. Faye (2010). Plasmodium falciparum GPI toxin: A common foe for man and mosquito. Acta Tropica 114, 162165.

N.T. Bech-Hansen , M.J. Naylor , T.A. Maybaum , R.L. Sparkes , B. Koop , D.G. Birch , A.A. Bergen , C.F. Prinsen , R.C. Polomeno , A. Gal , A.V. Drack , M.A. Musarella , S.G. Jacobson , R.S. Young & R.G. Weleber (2000). Mutations in NYX, encoding the leucine-rich proteoglycan nyctalopin, cause X-linked complete congenital stationary night blindness. Nature Genetics 26, 319323.

K.F. Chen , N. Peschel , R. Zavodska , H. Sehadova & R. Stanewsky (2011). QUASIMODO, a Novel GPI-anchored zona pellucida protein involved in light input to the Drosophila circadian clock. Current Biology 21, 719729.

N.J. Colley , E.K. Baker , M.A. Stamnes & C.S. Zuker (1991). The cyclophilin homolog ninaA is required in the secretory pathway. Cell 67, 255263.

N.J. Colley , J.A. Cassill , E.K. Baker & C.S. Zuker (1995). Defective intracellular transport is the molecular basis of rhodopsin-dependent dominant retinal degeneration. Proceedings of the National Academy of Sciences of the United States of America 92, 30703074.

J.M. Cordy , I. Hussain , C. Dingwall , N.M. Hooper & A.J. Turner (2003). Exclusively targeting beta-secretase to lipid rafts by GPI-anchor addition up-regulates beta-site processing of the amyloid precursor protein. Proceedings of the National Academy of Sciences of the United States of America 100, 1173511740.

S. David , E.J. Fry & R. Lopez-Vales (2008). Novel roles for Nogo receptor in inflammation and disease. Trends in Neurosciences 31, 221226.

A.L. Fabre , P. Orlean & C.H. Taron (2005). Saccharomyces cerevisiae Ybr004c and its human homologue are required for addition of the second mannose during glycosylphosphatidylinositol precursor assembly. The FEBS Journal 272, 11601168.

G.L. Fain , R. Hardie & S.B. Laughlin (2010). Phototransduction and the evolution of photoreceptors. Current Biology 20, R114R124.

C. Faivre-Sarrailh , S. Banerjee , J. Li , M. Hortsch , M. Laval & M.A. Bhat (2004). Drosophila contactin, a homolog of vertebrate contactin, is required for septate junction organization and paracellular barrier function. Development 131, 49314942.

N. Franceschini (1972). Pupil and pseudopupil in the compound eye of Drosophila. In Information Processing in the Visual Systems of Arthropods, ed. R. Wehner , pp. 7582. Berlin, Germany: Springer-Verlag.

N. Franceschini & K. Kirschfeld (1971). Pseudopupil phenomena in the Drosophila compound eye. Kybernetik 9, 159182.

M. Fujita & Y. Jigami (2008). Lipid remodeling of GPI-anchored proteins and its function. Biochimica et Biophysica Acta 1780, 410420.

S.J. Grimme , B.A. Westfall , J.M. Wiedman , C.H. Taron & P. Orlean (2001). The essential Smp3 protein is required for addition of the side-branching fourth mannose during assembly of yeast glycosylphosphatidylinositols. The Journal of Biological Chemistry 276, 2773127739.

R.C. Hardie & P. Raghu (2001). Visual transduction in Drosophila. Nature 413, 186193.

D. Hicks , D. John , N.Z. Makova , Z. Henderson , N.N. Nalivaeva & A.J. Turner (2011). Membrane targeting, shedding and protein interactions of brain acetylcholinesterase. Journal of Neurochemistry 116, 742746.

Y. Hirai-Fujita , M. Yamamoto-Hino , O. Kanie & S. Goto (2008). N-Glycosylation of the Drosophila neural protein Chaoptin is essential for its stability, cell surface transport and adhesive activity. FEBS Letters 582, 25722576.

J.Y. Kang , Y. Hong , H. Ashida , N. Shishioh , Y. Murakami , Y.S. Morita , Y. Maeda & T. Kinoshita (2005). PIG-V involved in transferring the second mannose in glycosylphosphatidylinositol. The Journal of Biological Chemistry 280, 94899497.

Y. Kanie , M. Yamamoto-Hino , Y. Karino , H. Yokozawa , S. Nishihara , R. Ueda , S. Goto & O. Kanie (2009). Insight into the regulation of glycan synthesis in Drosophila chaoptin based on mass spectrometry. PLoS One 4, e5434.

D. Karagogeos (2003). Neural GPI-anchored cell adhesion molecules. Frontiers in Bioscience 8, s1304s1320.

B. Katz & B. Minke (2009). Drosophila photoreceptors and signaling mechanisms. Frontiers in Cellular Neuroscience 3, 2.

S. Khandhadia , V. Cipriani , J.R. Yates & A.J. Lotery (2011). Age-related macular degeneration and the complement system. Immunobiology 217, 127146.

T. Kinoshita , M. Fujita & Y. Maeda (2008). Biosynthesis, remodelling and functions of mammalian GPI-anchored proteins: Recent progress. Journal of Biochemistry 144, 287294.

E.J. Koundakjian , D.M. Cowan , R.W. Hardy & A.H. Becker (2004). The Zuker collection: A resource for the analysis of autosomal gene function in Drosophila melanogaster. Genetics 167, 203206.

U.K. Laemmli (1970). Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227, 680685.

I. Levental , M. Grzybek & K. Simons (2010). Greasing their way: Lipid modifications determine protein association with membrane rafts. Biochemistry 49, 63056316.

C. Lopez-Pedrera , M.A. Aguirre , N. Barbarroja & M.J. Cuadrado (2010). Accelerated atherosclerosis in systemic lupus erythematosus: Role of proinflammatory cytokines and therapeutic approaches. Journal of Biomedicine & Biotechnology, pii: 607084, Epub 2010 Sep 26.

K.N. Ma , S.M. Cashman , J.H. Sweigard & R. Kumar-Singh (2010). Decay accelerating factor (CD55)-mediated attenuation of complement: Therapeutic implications for age-related macular degeneration. Investigative Ophthalmology & Visual Science 51, 67766783.

Y. Maeda & T. Kinoshita (2011). Structural remodeling, trafficking and functions of glycosylphosphatidylinositol-anchored proteins. Progress in Lipid Research 50, 411424.

S.E. Maxwell , S. Ramalingam , L.D. Gerber & S. Udenfriend (1995 b). Cleavage without anchor addition accompanies the processing of a nascent protein to its glycosylphosphatidylinositol-anchored form. Proceedings of the National Academy of Sciences of the United States of America 92, 15501554.

G. Maydan , I. Noyman , A. Har-Zahav , Z.B. Neriah , M. Pasmanik-Chor , A. Yeheskel , A. Albin-Kaplanski , I. Maya , N. Magal , E. Birk , A.J. Simon , A. Halevy , G. Rechavi , M. Shohat , R. Straussberg & L. Basel-Vanagaite (2011). Multiple congenital anomalies-hypotonia-seizures syndrome is caused by a mutation in PIGN. Journal of Medical Genetics 48, 383389.

K. Nagamune , K. Ohishi , H. Ashida , Y. Hong , J. Hino , K. Kangawa , N. Inoue , Y. Maeda & T. Kinoshita (2003). GPI transamidase of Trypanosoma brucei has two previously uncharacterized (trypanosomatid transamidase 1 and 2) and three common subunits. Proceedings of the National Academy of Sciences of the United States of America 100, 1068210687.

E. O’Connor , B. Eisenhaber , J. Dalley , T. Wang , C. Missen , N. Bulleid , P.N. Bishop & D. Trump (2005). Species specific membrane anchoring of nyctalopin, a small leucine-rich repeat protein. Human Molecular Genetics 14, 18771887.

R. Oriol , I. Martinez-Duncker , I. Chantret , R. Mollicone & P. Codogno (2002). Common origin and evolution of glycosyltransferases using Dol-P-monosaccharides as donor substrate. Molecular Biology and Evolution 19, 14511463.

P. Orlean & A.K. Menon (2007). GPI anchoring of protein in yeast and mammalian cells, or: How we learned to stop worrying and love glycophospholipids. Journal of Lipid Research 48, 9931011.

N. Ortonne , C. Ram-Wolff , J. Giustiniani , A. Marie-Cardine , M. Bagot , S. Mecheri & A. Bensussan (2011). Human and mouse mast cells express and secrete the GPI-anchored isoform of CD160. The Journal of Investigative Dermatology 131, 916924.

S. Pang , P. Urquhart & N.M. Hooper (2004). N-glycans, not the GPI anchor, mediate the apical targeting of a naturally glycosylated, GPI-anchored protein in polarised epithelial cells. Journal of Cell Science 117, 50795086.

M.G. Paulick & C.R. Bertozzi (2008). The glycosylphosphatidylinositol anchor: A complex membrane-anchoring structure for proteins. Biochemistry 47, 69917000.

K. Ramo , S.M. Cashman & R. Kumar-Singh (2008). Evaluation of adenovirus-delivered human CD59 as a potential therapy for AMD in a model of human membrane attack complex formation on murine RPE. Investigative Ophthalmology & Visual Science 49, 41264136.

E.E. Rosenbaum , R.C. Hardie & N.J. Colley (2006). Calnexin is essential for rhodopsin maturation, Ca2+ regulation, and photoreceptor cell survival. Neuron 49, 229241.

P. Ruiz , D. Weppler , P. Tryphonopoulos , S. Nishida , J. Moon , T. Kato , G. Selvaggi , D. Levi , J. Madariaga , J. DelaGarza , S. Tuteja , M. Garcia & A. Tzakis (2006). CD55 and CD59 deficiency in transplant patient populations: Possible association with paroxysmal nocturnal hemoglobinuria-like symptoms in Campath-treated patients. Transplantation Proceedings 38, 17501752.

F.J. Sharom & M.T. Lehto (2002). Glycosylphosphatidylinositol-anchored proteins: Structure, function, and cleavage by phosphatidylinositol-specific phospholipase C. Biochemistry and Cell Biology 80, 535549.

T.D. Spurway , J.A. Dalley , S. High & N.J. Bulleid (2001). Early events in glycosylphosphatidylinositol anchor addition. substrate proteins associate with the transamidase subunit gpi8p. The Journal of Biological Chemistry 276, 1597515982.

A. Tiede , I. Bastisch , J. Schubert , P. Orlean & R.E. Schmidt (1999). Biosynthesis of glycosylphosphatidylinositols in mammals and unicellular microbes. Biological Chemistry 380, 503523.

O. Vagin , J.A. Kraut & G. Sachs (2009). Role of N-glycosylation in trafficking of apical membrane proteins in epithelia. American Journal of Physiology. Renal Physiology 296, F459F469.

D. Van Vactor Jr, D.E. Krantz , R. Reinke & S.L. Zipursky (1988). Analysis of mutants in Chaoptin, a photoreceptor cell-specific glycoprotein in Drosophila, reveals its role in cellular morphogenesis. Cell 52, 281290.

Y. Varma & T. Hendrickson (2010). Methods to study GPI anchoring of proteins. Chembiochem 11, 623636.

R. Veerhuis (2011). Histological and direct evidence for the role of complement in the neuroinflammation of AD. Current Alzheimer Research 8, 3458.

R. Veerhuis , H.M. Nielsen & A.J. Tenner (2011). Complement in the brain. Molecular Immunology 48, 15921603.

K.S. Vetrivel , A. Barman , Y. Chen , P.D. Nguyen , S.L. Wagner , R. Prabhakar & G. Thinakaran (2011). Loss of cleavage at beta’-site contributes to apparent increase in beta-amyloid peptide (Abeta) secretion by beta-secretase (BACE1)-glycosylphosphatidylinositol (GPI) processing of amyloid precursor protein. The Journal of Biological Chemistry 286, 2616626177.

T. Wang & C. Montell (2007). Phototransduction and retinal degeneration in Drosophila. Pflügers Archiv 454, 821847.

K.W. Yau & R.C. Hardie (2009). Phototransduction motifs and variations. Cell 139, 246264.

C. Zeitz , H. Scherthan , S. Freier , S. Feil , V. Suckow , S. Schweiger & W. Berger (2003). NYX (nyctalopin on chromosome X), the gene mutated in congenital stationary night blindness, encodes a cell surface protein. Investigative Ophthalmology & Visual Science 44, 41844191.

J. Zhang , C. Gerhardinger & M. Lorenzi (2002). Early complement activation and decreased levels of glycosylphosphatidylinositol-anchored complement inhibitors in human and experimental diabetic retinopathy. Diabetes 51, 34993504.

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Visual Neuroscience
  • ISSN: 0952-5238
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