In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import

Particle and Fibre Toxicology

Timothy P Yoshino¹  Xiao-Jun Wu¹  Tavis K Anderson³  Nathan A Peterson² 

[1] Current address: Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, 2115 Observatory Drive, Madison, WI 53706, USA;Current address: Department of Entomology, College of Agricultural and Life Sciences, University of Wisconsin, 1630 Linden Drive, Madison, WI 53706, USA;Current address: Virus and Prion Research Unit, National Animal Disease Center, USDA, Agricultural Research Service, 1920 Dayton Ave, Ames, IA 50010, USA

关键词: Sporocyst;    Miracidium;    Fucosyltransferase;    GDP-L-fucose transport;    GDP-L-fucose synthesis;    Fucosylation;    Fucose;    Schistosome;    Schistosoma mansoni;   

Others  :  1226163
DOI  :  10.1186/1756-3305-6-201

 received in 2013-01-23, accepted in 2013-06-15,  发布年份 2013

【 摘 要 】

Background

Carbohydrate structures of surface-expressed and secreted/excreted glycoconjugates of the human blood fluke Schistosoma mansoni are key determinants that mediate host-parasite interactions in both snail and mammalian hosts. Fucose is a major constituent of these immunologically important glycans, and recent studies have sought to characterize fucosylation-associated enzymes, including the Golgi-localized fucosyltransferases that catalyze the transfer of L-fucose from a GDP-L-fucose donor to an oligosaccharide acceptor. Importantly, GDP-L-fucose is the only nucleotide-sugar donor used by fucosyltransferases and its availability represents a bottleneck in fucosyl-glycotope expression.

Methods

A homology-based genome-wide bioinformatics approach was used to identify and molecularly characterize the enzymes that contribute to GDP-L-fucose synthesis and Golgi import in S. mansoni. Putative functions were further investigated through molecular phylogenetic and immunocytochemical analyses.

Results

We identified homologs of GDP-D-mannose-4,6-dehydratase (GMD) and GDP-4-keto-6-deoxy-D-mannose-3,5-epimerase-4-reductase (GMER), which constitute a de novo pathway for GDP-L-fucose synthesis, in addition to a GDP-L-fucose transporter (GFT) that putatively imports cytosolic GDP-L-fucose into the Golgi. In silico primary sequence analyses identified characteristic Rossman loop and short-chain dehydrogenase/reductase motifs in GMD and GMER as well as 10 transmembrane domains in GFT. All genes are alternatively spliced, generating variants of unknown function. Observed quantitative differences in steady-state transcript levels between miracidia and primary sporocysts may contribute to differential glycotope expression in early larval development. Additionally, analyses of protein expression suggest the occurrence of cytosolic GMD and GMER in the ciliated epidermal plates and tegument of miracidia and primary sporocysts, respectively, which is consistent with previous localization of highly fucosylated glycotopes.

Conclusions

This study is the first to identify and characterize three key genes that are putatively involved in the synthesis and Golgi import of GDP-L-fucose in S. mansoni and provides fundamental information regarding their genomic organization, genetic variation, molecular phylogenetics, and developmental expression in intramolluscan larval stages.

【 授权许可】

   
2013 Peterson et al.; licensee BioMed Central Ltd.

【 图 表 】

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