Ectophosphodiesterase/nucleotide phosphohydrolase (Enpp) nucleotidases: cloning, conservation and developmental restriction
Karine Masse, Surinder Bhamra, Gavin Allsop, Nicholas Dale, Elizabeth A. Jones
2010
International Journal of Developmental Biology
Ectonucleotidase proteins occupy a central role in purine signalling regulation by sequentially hydrolysing ATP to ADP and to adenosine. The ENPP ( or PDNP) gene family, which encodes ectophosphodiesterase/nucleotide phosphohydrolases, is a subfamily of these enzymes, which consists of 7 members in mammals. These proteins catalyse the generation of bioactive lipids, placing the ENPP enzymes as key regulators of major physiological signalling pathways and also important players in several
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... gical conditions. Here we report the cloning of all the members, except enpp5, of the enpp family in Xenopus laevis and tropicalis. Phylogenetic analyses demonstrate the high level of conservation of these proteins between amphibian and other vertebrate species. During development and in the adult frog, each gene displays a distinct specific expression pattern, suggesting potentially different functions for these proteins during amphibian embryogenesis. This is the first complete developmental analysis of gene expression of this gene family in vertebrates. Results Cloning of the different enpp genes All the members of the enpp family, except enpp5, were cloned using different strategies (Supplementary Table 1A ). TBLASTN search of the X. laevis and tropicalis EST databases on the NCBI website allowed the identification of full length IMAGE clones or EST Unigenes coding for enpp2, enpp4, enpp6 and enpp7 (Accession numbers given in Supplementary Table 1B) . When necessary, alignments of these sequences were performed to generate the consensus sequence of the cDNA. BLAST search on ESTs databases only allowed the cloning of partial sequences for Xenopus enpp1 and enpp3 cDNAs. The remaining sequences were identified by BLAST on the genomic X. tropicalis databases (JGI website) and the deduced cDNA sequences corrected by reference to the human sequence according to the Breathnach and Chambon law (Breathnach and Chambon, 1981) . When needed, RT-PCRs were performed on X. laevis and tropicalis embryonic and adult tissues to amplify the missing sequences. Protein and phylogenetic analysis of the enpp family Consensus protein sequence deduced from conceptual translation of the cDNA sequences are given in Figs. 1 and 2 and each Xenopus enpp protein displays the characteristics features of this
doi:10.1387/ijdb.092879km
pmid:19598106
fatcat:3humm3qvvzafvmpxhuujusxtte