Cloning And Sequencing Of Two CDNAs Encoding The Catalytic Beta Subunit Of Protein Phosphatase 1 And Metalloprotease Of Flatfish Turbot Scophthalmus Maximus By RACE Technique

Reversible protein phosphorylation, catalysed by protein kinases and phosphatases, is an important and versatile mechanism by which eukaryotic cells regulate almost all the signaling processes. Protein phosphatase 1 (PP1) is the first and well characterized member of the protein serine/threonine phosphatase family. In the present study, a full-length cDNA encoding the beta isoform of the catalytic subunit of protein phosphatase 1(PP1cb), was for the first time isolated and sequenced from the skin tissue of flatfish turbot Scophthalmus maximus, designated SmPP1cb, by Rapid amplification of cDNA ends (RACE) technique. The cDNA sequence of SmPP1cb we obtained contains a 984 bp open reading frame (ORF), flanked by a complete 39 bp 5'untranslated region and 462 bp 3'untranslated region. The ORF encodes a putative 327 amino acid protein with a characteristic highly acidic N-terminal region, Met-Ala-Glu-Gly-Glu-Leu-Asp-Val-Asp. And its calculated molecular mass is 37,193 Da and pI 5.8. Sequence analysis indicated that, SmPP1cb is extremely conserved in both amino acid and nucleotide acid levels compared with the PP1cb of other vertebrates and invertebrates, and contains a favorable 5'UTR , with a different context surrounding ATG codon from mammalian, ensuring its efficient translation initiation in turbot and a highly...