Molecular cloning and characterization of UDP-glucose dehydrogenase from the amphibian Xenopus laevis and its involvement in hyaluronan synthesis.
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.
Vigetti
Davide
D
Dipartimento di Scienze Biomediche Sperimentali e Cliniche, Università degli Studi dell'Insubria, via J. H. Dunant 5, 21100 Varese, Italy.
Ori
Michela
M
Viola
Manuela
M
Genasetti
Anna
A
Karousou
Eugenia
E
Rizzi
Manuela
M
Pallotti
Francesco
F
Nardi
Irma
I
Hascall
Vincent C
VC
De Luca
Giancarlo
G
Passi
Alberto
A
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
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