title: Molecular cloning and characterization of UDP-glucose dehydrogenase from the amphibian Xenopus laevis and its involvement in hyaluronan synthesis.
abstract: UDP-glucose dehydrogenase (UGDH) supplies the cell with UDP-glucuronic acid (UDP-GlcUA), a precursor of glycosaminoglycan and proteoglycan synthesis. Here we reported the cloning and the characterization of the UGDH from the amphibian Xenopus laevis that is one of the model organisms for developmental biology. We found that X. laevis UGDH (xUGDH) maintained a very high degree of similarity with other known UGDH sequences both at the genomic and the protein levels. Also its kinetic parameters are similar to those of UGDH from other species. During X. laevis development, UDGH is always expressed but clearly increases its mRNA levels at the tail bud stage (i.e. 30 h post-fertilization). This result fits well with our previous observation that hyaluronan, a glycosaminoglycan that is synthesized using UDP-GlcUA and UDP-N-acetylglucosamine, is abundantly detected at this developmental stage. The expression of UGDH was found to be related to hyaluronan synthesis. In human smooth muscle cells the overexpression of xUGDH or endogenous abrogation of UGDH modulated hyaluronan synthesis specifically. Our findings were confirmed by in vivo experiments where the silencing of xUGDH in X. laevis embryos decreased glycosaminoglycan synthesis causing severe embryonic malformations because of a defective gastrulation process.
authors.affiliation: Dipartimento di Scienze Biomediche Sperimentali e Cliniche, Università degli Studi dell'Insubria, via J. H. Dunant 5, 21100 Varese, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Vigetti
authors.forename: Michela
authors.initials: M
authors.lastname: Ori
authors.forename: Manuela
authors.initials: M
authors.lastname: Viola
authors.forename: Anna
authors.initials: A
authors.lastname: Genasetti
authors.forename: Eugenia
authors.initials: E
authors.lastname: Karousou
authors.forename: Manuela
authors.initials: M
authors.lastname: Rizzi
authors.forename: Francesco
authors.initials: F
authors.lastname: Pallotti
authors.forename: Irma
authors.initials: I
authors.lastname: Nardi
authors.forename: Vincent C
authors.initials: VC
authors.lastname: Hascall
authors.forename: Giancarlo
authors.initials: G
authors.lastname: De Luca
authors.forename: Alberto
authors.initials: A
authors.lastname: Passi
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017077:Culture Media, Conditioned / Q000494:pharmacology; D017931:DNA Primers / Q000737:chemistry; D004187:Disaccharides / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D005091:Exons; D020224:Expressed Sequence Tags; D005775:Gastrula / Q000378:metabolism; D015870:Gene Expression; D005786:Gene Expression Regulation; D018507:Gene Expression Regulation, Developmental; D020868:Gene Silencing; D016678:Genome; D006025:Glycosaminoglycans / Q000737:chemistry; D006801:Humans; D006820:Hyaluronic Acid / Q000096:biosynthesis* / Q000737:chemistry; D007438:Introns; D007700:Kinetics; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014533:Uridine Diphosphate Glucose Dehydrogenase / Q000737:chemistry* / Q000235:genetics*; D014535:Uridine Diphosphate Glucuronic Acid / Q000737:chemistry; D014981:Xenopus / Q000378:metabolism; D014982:Xenopus laevis
chemical_list: D017077:Culture Media, Conditioned; D017931:DNA Primers; D004187:Disaccharides; D006025:Glycosaminoglycans; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D014535:Uridine Diphosphate Glucuronic Acid; D006820:Hyaluronic Acid; D014533:Uridine Diphosphate Glucose Dehydrogenase