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J. Biol. Chem., Vol. 281, Issue 12, 8254-8263, March 24, 2006

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Molecular Cloning and Characterization of UDP-glucose Dehydrogenase from the Amphibian Xenopus laevis and Its Involvement in Hyaluronan Synthesis^*

Davide Vigetti, Michela Ori, Manuela Viola, Anna Genasetti, Eugenia Karousou, Manuela Rizzi, Francesco Pallotti, Irma Nardi, Vincent C. Hascall^¶, Giancarlo De Luca, and Alberto Passi¹

From the Dipartimento di Scienze Biomediche Sperimentali e Cliniche, Università degli Studi dell'Insubria, via J. H. Dunant 5, 21100 Varese, Italy, Laboratori di Biologia Cellulare e dello Sviluppo, Dipartimento di Fisiologia e Biochimica, Università di Pisa, via Carducci 13, Ghezzano, 56010 Pisa, Italy, and the ^¶Department of Biomedical Engineering and Orthopaedic Research Center/ND20, The Cleveland Clinic Foundation, Cleveland, Ohio 44195

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.

Received for publication, August 3, 2005 , and in revised form, December 22, 2005.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AY762616 [GenBank] .

^* This work was supported by MIUR COFIN Project "Analysis of the Hyaluronan-Proteoglycan-Hyaluronan Receptor System in Embryonic Development and Disease" (to D. V.) and Consorzio Interuniversitario Biotecnologie (to A. P.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Dip. di Scienze Biomediche Sperimentali e Cliniche, Università degli Studi dell'Insubria, via J. H. Dunant 5, 21100 Varese, Italy. Tel.: 39-0332-217142; Fax: 39-0332-217119; E-mail: alberto.passi{at}uninsubria.it.

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