Cloning and sequence analysis of a full-length cDNA of SmPPlcb encoding turbot protein phosphatase 1 beta catalytic subunit

摘要

Reversible protein phosphorylation,catalyzed 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 l(PPlcb),was for the first time isolated and sequenced from the skin tissue of flatfish turbot Scophthalmus maximus,designated SmPPlcb,by the rapid amplification of cDNA ends (RACE) technique.The cDNA sequence of SmPPlcb 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,and the N-terminal section of this protein is highly acidic,Met-Ala-Glu-Gly-Glu-Leu-Asp-Val-Asp,a common feature for PP1 catalytic subunit but absent in protein phosphatase 2B (PP2B).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 its Kozak motif contained in the 5'UTR around ATG start codon is GXXAXXGXXATGG which is different from mammalian in two positions A~(-6) and G~(-3),indicating the possibility of different initiation of translation in turbot,and also the 3'UTR of SmPP1cb is highly diverse in the sequence similarity and length compared with other animals,especially zebrafish.The cloning and sequencing of SmPP1cb gene lays a good foundation for the future work on the biological functions of PP1 in the flatfish turbot.