[en] In holometabolous insects the transition from larva to adult requires a complete body reorganization
and relies on N-glycosylated proteins. N-glycosylation is an important posttranslational modification
that influences protein activity but its impact on the metamorphosis has not been studied yet. Here
we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the
involvement of the protein N-glycosylation pathway in metamorphosis. The transcript levels for genes
encoding N-glycan processing enzymes increased during later developmental stages and, in turn,
transition from larva to adult coincided with an enrichment of more extensively modified paucimannose
glycans, including fucosylated ones. Blockage of N-glycan attachment resulted in larval mortality, while
RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva
to pupa transition. Additionally, simultaneous knockdown of multiple genes responsible for N-glycan
processing towards paucimannose structures revealed their novel roles in pupal appendage formation
and adult eclosion. Our findings revealed that, next to hormonal control, insect post-embryonic
development and metamorphosis depend on protein N-glycan attachment and efficient N-glycan
processing. Consequently, disruption of these processes could be an effective new approach for insect
control.
Disciplines :
Entomology & pest control
Author, co-author :
Walski, Tomasz; Universiteit Gent - Ugent > Department of Crop Protection > Laboratory of Agrozoology
Van Damme, Els; Universiteit Gent - Ugent > Department of Molecular Biotechnology > Laboratory of Biochemistry and Glycobiology
Smargiasso, Nicolas ; Université de Liège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Christiaens, Olivier; Universiteit Gent - Ugent > Department of Crop Protection > Laboratory of Agrozoology
De Pauw, Edwin ; Université de Liège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Smagghe, Guy; Universiteit Gent - Ugent > Department of Crop Protection > Laboratory of Agrozoology
Language :
English
Title :
Protein N-glycosylation and N-glycan trimming are required for postembryonic development of the pest beetle Tribolium castaneum
Stanley, P., Schachter, H., Taniguchi, N. N-glycans. In: Essentials of Glycobiology (eds Varki A CR, Esko JD et al. ). 2nd edn. Cold Spring Harbor Laboratory Press (2009).
H elenius A., Aebi, M. Roles of N-linked glycans in the endoplasmic reticulum. Annu. Rev. Biochem. 73, 1019-1049 (2004).
Moremen, K. W., Tiemeyer, M., Nairn, A. V. Vertebrate protein glycosylation: diversity, synthesis and function. Nat. Rev. Mol. Cell Biol. 13, 448-462 (2012).
Kurz, S. et al. Targeted release and fractionation reveal glucuronylated and sulphated N-and O-glycans in larvae of dipteran insects. J. Prot. 126, 172-188 (2015).
Kimura, Y. et al. Tumor antigen occurs in N-glycan of royal jelly glycoproteins: Honeybee cells synthesize T-antigen unit in N-glycan moiety. Biosci. Biotech. Bioch. 70, 2583-2587 (2006).
Kurz, S., King, J. G., Dinglasan, R. R., Paschinger, K., Wilson, I. B. The fucomic potential of mosquitoes: fucosylated N-glycan epitopes and their cognate fucosyltransferases. Insect Biochem. Mol. Biol. 68, 52-63 (2016).
Aoki, K., Tiemeyer, M. Chapter Fourteen-The Glycomics of Glycan Glucuronylation in Drosophila melanogaster. Methods Enzymol. 480, 297-321 (2010).
Rendic, D., Wilson, I. B. H., Paschinger, K. The glycosylation capacity of insect cells. Croat. Chem. Acta 81, 7-21 (2008).
Sekine, S. U. et al. Meigo governs dendrite targeting specificity by modulating ephrin level and N-glycosylation. Nat. Neurosci. 16, 683-691 (2013).
Koles, K. et al. Identification of N-glycosylated proteins from the central nervous system of Drosophila melanogaster. Glycobiology 17, 1388-1403 (2007).
Sarkar, M., Iliadi, K. G., Leventis, P. A., Schachter, H., Boulianne, G. L. Neuronal expression of Mgat1 rescues the shortened life span of Drosophila Mgat11 null mutants and increases life span. Proc. Natl. Acad. Sci. USA 107, 9677-9682 (2010).
Mortimer, N. T., Kacsoh, B. Z., Keebaugh, E. S., Schlenke, T. A. Mgat1-dependent N-glycosylation of membrane components primes Drosophila melanogaster blood cells for the cellular encapsulation response. PLOS Pathog. 8, e1002819 (2012).
Gui, Z. Z. et al. Functional role of aspartic proteinase cathepsin D in insect metamorphosis. BMC Dev. Biol. 6, 49 (2006).
Zielinska, D. F., Gnad, F., Schropp, K., Winiewski, J. R., Mann, M. Mapping N-glycosylation sites across seven evolutionarily distant species reveals a divergent substrate proteome despite a common core machinery. Mol. Cell 46, 542-548 (2012).
Vandenborre, G. et al. Diversity in protein glycosylation among insect species. PLOS ONE 6, e16682 (2011).
Zhang, J., Lu, A., Kong, L., Zhang, Q., Ling, E. Functional analysis of insect molting fluid proteins on the protection and regulation of ecdysis. J. Biol. Chem. 289, 35891-35906 (2014).
Jindra, M., Palli, S. R., Riddiford, L. M. The juvenile hormone signaling pathway in insect development. Annu. Rev. Entomol. 58, 181-204 (2013).
Truman, J. W., Riddiford, L. M. Endocrine insights into the evolution of metamorphosis in insects. Annu. Rev. Entomol. 47, 467-500 (2002).
Linz, D. M., Tomoyasu, Y. RNAi screening of developmental toolkit genes: a search for novel wing genes in the red flour beetle, Tribolium castaneum. Dev. Genes Evol. 225, 11-22 (2015).
Tunaz, H., Uygun, N. Insect growth regulators for insect pest control. Turk. J. Agric. For. 28, 377-387 (2004).
Wilson, T. G. The molecular site of action of juvenile hormone and juvenile hormone insecticides during metamorphosis: how these compounds kill insects. J. Insect Physiol. 50, 111-121 (2004).
Nemcovicova, I. et al. Characterisation of class I and II alpha-mannosidases from Drosophila melanogaster. Glycoconj. J. 30, 899-909 (2013).
Dragosits, M., Yan, S., Razzazi-Fazeli, E., Wilson, I. B., Rendic, D. Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. Glycobiology 25, 448-464 (2015).
Vadaie, N., Jarvis, D. L. Molecular cloning and functional characterization of a Lepidopteran insect beta4-Nacetylgalactosaminyltransferase with broad substrate specificity, a functional role in glycoprotein biosynthesis, and a potential functional role in glycolipid biosynthesis. J. Biol. Chem. 279, 33501-33518 (2004).
Rosenbaum, E. E., Vasiljevic, E., Brehm, K. S., Colley, N. J. Mutations in four glycosyl hydrolases reveal a highly coordinated pathway for rhodopsin biosynthesis and N-glycan trimming in Drosophila melanogaster. PLOS Genet. 10, e1004349 (2014).
Graveley, B. R. et al. The developmental transcriptome of Drosophila melanogaster. Nature 471, 473-479 (2011).
Gelbart, W. M., Emmert, D. B. FlyBase High Throughput Expression Pattern Data. (2013). Available from: http://flybase. org. Accessed on: 12/09/2015.
Leonard, R. et al. The Drosophila fused lobes gene encodes an N-acetylglucosaminidase involved in N-glycan processing. J. Biol. Chem. 281, 4867-4875 (2006).
Mabashi-Asazuma, H. et al. Targeted Glycoengineering Extends the Protein N-glycosylation Pathway in the Silkworm Silk Gland. Insect Biochem. Mol. Biol. 65, 20-27 (2015).
Cipollo, J. F., Awad, A. M., Costello, C. E., Hirschberg, C. B. N-Glycans of Caenorhabditis elegans are specific to developmental stages. J. Biol. Chem. 280, 26063-26072 (2005).
Fabini, G., Freilinger, A., Altmann, F., Wilson, I. B. Identification of core alpha 1, 3-fucosylated glycans and cloning of the requisite fucosyltransferase cDNA from Drosophila melanogaster. Potential basis of the neural anti-horseadish peroxidase epitope. J. Biol. Chem. 276, 28058-28067 (2001).
Aoki, K. et al. Dynamic developmental elaboration of N-linked glycan complexity in the Drosophila melanogaster embryo. J. Biol. Chem. 282, 9127-9142 (2007).
Rendi, D. et al. Modulation of neural carbohydrate epitope expression in Drosophila melanogaster cells. J. Biol. Chem. 281, 3343-3353 (2006).
Repnikova, E. et al. Sialyltransferase regulates nervous system function in Drosophila. J. Neurosci. 30, 6466-6476 (2010).
Shrimal, S., Trueman, S. F., Gilmore, R. Extreme C-terminal sites are posttranslocationally glycosylated by the STT3B isoform of the OST. J. Cell Biol. 201, 81-95 (2013).
Sato, T. et al. STT3B-dependent posttranslational N-glycosylation as a surveillance system for secretory protein. Mol. Cell 47, 99-110 (2012).
Spradling, A. C. et al. The Berkeley Drosophila Genome Project gene disruption project: single P-element insertions mutating 25% of vital Drosophila genes. Genetics 153, 135-177 (1999).
Ruiz-Canada, C., Kelleher, D. J., Gilmore, R. Cotranslational and posttranslational N-glycosylation of polypeptides by distinct mammalian OST isoforms. Cell 136, 272-283 (2009).
Sanjay, A., Fu, J., Kreibich, G. DAD1 is required for the function and the structural integrity of the oligosaccharyltransferase complex. J. Biol. Chem. 273, 26094-26099 (1998).
Ditz, J. et al. iBeetle-Base: a database for RNAi phenotypes in the red flour beetle Tribolium castaneum. Nucleic Acids Res. gku1054 (2014).
Katoh, T. et al. Deficiency of alpha-glucosidase I alters glycoprotein glycosylation and lifespan in Caenorhabditis elegans. Glycobiology 23, 1142-1151 (2013).
Mummery-Widmer, J. L. et al. Genome-wide analysis of Notch signalling in Drosophila by transgenic RNAi. Nature 458, 987-992 (2009).
Kerscher, S., Albert, S., Wucherpfennig, D., Heisenberg, M., Schneuwly, S. Molecular and genetic analysis of the Drosophila mas-1 (mannosidase-1) gene which encodes a glycoprotein processing 1, 2-mannosidase. Dev. Biol. 168, 613-626 (1995).
Roberts, D. B., Mulvany, W. J., Dwek, R. A., Rudd, P. M. Mutant analysis reveals an alternative pathway for N-linked glycosylation in Drosophila melanogaster. Eur. J. Biochem. 253, 494-498 (1998).
Kawar, Z., Karaveg, K., Moremen, K. W., Jarvis, D. L. Insect cells encode a class II -mannosidase with unique properties. J. Biol. Chem. 276, 16335-16340 (2001).
Oh-eda, M. et al. Overexpression of the Golgi-localized enzyme -mannosidase IIx in Chinese hamster ovary cells results in the conversion of hexamannosyl-N-acetylchitobiose to tetramannosyl-N-acetylchitobiose in the N-glycan-processing pathway. Eur. J. Biochem. 268, 1280-1288 (2001).
Altmann, F., Kornfeld, G., Dalik, T., Staudacher, E., Glsl, J. Processing of asparagine-linked oligosaccharides in insect cells. N-Acetylglucosaminyltransferase I and II activities in cultured lepidopteran cells. Glycobiology 3, 619-625 (1993).
Schachter, H. Mgat1-dependent N-glycans are essential for the normal development of both vertebrate and invertebrate metazoans. Semin. Cell Dev. Biol. 21, 609-615 (2010).
Zhang, W. et al. Synthesis of paucimannose N-glycans by Caenorhabditis elegans requires prior actions of UDP-N-acetyl-Dglucosamine: alpha-3-D-mannoside beta1, 2-N-acetylglucosaminyltransferase I, alpha3, 6-mannosidase II and a specific membrane-bound beta-N-acetylglucosaminidase. Biochem. J 372, 53-64 (2003).
Paschinger, K., Staudacher, E., Stemmer, U., Fabini, G., Wilson, I. B. H. Fucosyltransferase substrate specificity and the order of fucosylation in invertebrates. Glycobiology 15, 463-474 (2005).
Kawar, Z., Romero, P. A., Herscovics, A., Jarvis, D. L. N-Glycan processing by a lepidopteran insect 1, 2-mannosidase. Glycobiology 10, 347-355 (2000).
Benyair, R., Ogen-Shtern, N., Lederkremer, G. Z. Glycan regulation of ER-associated degradation through compartmentalization. Semin. Cell Dev. Biol. 41, 99-109 (2015).
Lorenzen, M. et al. piggyBacmediated germline transformation in the beetle Tribolium castaneum. Insect Mol. Biol. 12, 433-440 (2003).
Yamamoto-Hino, M. et al. Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing. Genes Cells 20, 521-542 (2015).
Baycin-Hizal, D. et al. GlycoFly: a database of Drosophila N-linked glycoproteins identified using SPEG-MS techniques. J. Prot. Res. 10, 2777-2784 (2011).
Marada, S. et al. Functional Divergence in the Role of N-Linked Glycosylation in Smoothened Signaling. PLOS Genet. 11, e1005473 (2015).
Shingleton, A. W., Das, J., Vinicius, L., Stern, D. L. The temporal requirements for insulin signaling during development in Drosophila. PLOS Biol. 3, e289 (2005).
Broehan, G. et al. Chymotrypsin-like peptidases from Tribolium castaneum: A role in molting revealed by RNA interference. Insect Biochem. Mol. Biol. 40, 274-283 (2010).
Zhu, Q., Arakane, Y., Beeman, R. W., Kramer, K. J., Muthukrishnan, S. Functional specialization among insect chitinase family genes revealed by RNA interference. Proc. Natl. Acad. Sci. USA 105, 6650-6655 (2008).
Arakane, Y. et al. The Tribolium chitin synthase genes TcCHS1 and TcCHS2 are specialized for synthesis of epidermal cuticle and midgut peritrophic matrix. Insect Mol. Biol. 14, 453-463 (2005).
Domguez-Gimez, P., Brown, N. H., Mart-Bermudo, M. D. Integrin-ECM interactions regulate the changes in cell shape driving the morphogenesis of the Drosophila wing epithelium. J. Cell Sci. 120, 1061-1071 (2007).
Pesch, Y.-Y., Riedel, D., Patil, K. R., Loch, G., Behr, M. Chitinases and Imaginal disc growth factors organize the extracellular matrix formation at barrier tissues in insects. Sci. Rep. 6, 18340, doi:10. 1038/srep18340 (2016).
Hsu, T.-A. et al. Differential N-glycan patterns of secreted and intracellular IgG produced in Trichoplusia ni cells. J. Biol. Chem. 272, 9062-9070 (1997).
Yamamoto, A. et al. Neurogenetic networks for startle-induced locomotion in Drosophila melanogaster. Proc. Natl. Acad. Sci. USA 105, 12393-12398 (2008).
Beck, E. S. et al. Regulation of Fasciclin II and synaptic terminal development by the splicing factor beag. J. Neurosci. 32, 7058-7073 (2012).
Haines, N., Stewart, B. A. Functional Roles for 1, 4-N-Acetlygalactosaminyltransferase-A in Drosophila Larval Neurons and Muscles. Genetics 175, 671-679 (2007).
Kanie, Y. et al. Insight into the regulation of glycan synthesis in Drosophila chaoptin based on mass spectrometry. PLOS ONE, 4, e5434 (2009).
Hang, I. et al. Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. Glycobiology 25, 1335-1349 (2015).
Tamura, K. et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28, 2731-2739 (2011).
Ye, J. et al. Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics 13, 134 (2012).
Kang, P., Mechref, Y., Novotny, M. V. High-throughput solid-phase permethylation of glycans prior to mass spectrometry. Rapid Commun. Mass Spectrom. 22, 721-734 (2008).
Ceroni, A. et al. GlycoWorkbench: a tool for the computer-assisted annotation of mass spectra of glycans. J. Prot. Res. 7, 1650-1659 (2008).
Horn, T., Boutros, M. E-RNAi: a web application for the multi-species design of RNAi reagents-2010 update. Nucleic Acids Res. gkq317 (2010).
Clark-Hachtel, C. M., Linz, D. M., Tomoyasu, Y. Insights into insect wing origin provided by functional analysis of vestigial in the red flour beetle, Tribolium castaneum. Proc. Natl. Acad. Sci. USA 110, 16951-16956 (2013).
Arakane, Y., Muthukrishnan, S., Beeman, R. W., Kanost, M. R., Kramer, K. J. Laccase 2 is the phenoloxidase gene required for beetle cuticle tanning. Proc. Natl. Acad. Sci. USA 102, 11337-11342 (2005).
Schindelin, J. et al. Fiji: An open-source platform for biological-image analysis. Nat. Meth. 9, 676-682 (2012).
Lowe, D. G. Distinctive image features from scale-invariant keypoints. Int. J. Comput. Vis. 60, 91-110 (2004).
Aguet, F., Van De Ville, D., Unser, M. Model-based 2. 5-D deconvolution for extended depth of field in brightfield microscopy. IEEE Trans. Image Process. 17, 1144-1153 (2008).