Molecular characterization and expression analysis of five different elongation factor 1 alpha genes in the flatfish Senegalese sole(Solea senegalensis Kaup):Differential gene expression and thyroid hormones dependence during metamorphosis

摘要

Background:Eukaryotic elongation factor 1 alpha(eEF1A)is one of the four subunits composing eukaryotic translation elongation factor 1.It catalyzes the binding of aminoacyl-tRNA to the A-site of the ribosome in a GTP-dependent manner during protein synthesis,although it also seems to play a role in other non-translational processes.Currently,little information is still available about its expression profile and regulation during flatfish metamorphosis.With regard to this,Senegalese sole(Solea senegalensis)is a commercially important flatfish in which eEF1A gene remains to be characterized. Results:The development of large-scale genomics of Senegalese sole has facilitated the identification of five different eEF1A genes,referred to as SseEF1A1,SseEF1A2,SseEF1A3,SseEF1A4,and Sse42Sp50.Main characteristics and sequence identities with other fish and mammalian eEF1As are described.Phylogenetic and tissue expression analyses allowed for the identification of SseEF1A1 and SseEF1A2 as the Senegalese sole counterparts of mammalian eEF1A1 and eEF1A2,respectively,and of Sse42Sp50 as the ortholog of Xenopus laevis and teleost 42Sp50 gene.The other two elongation factors,SseEF1A3 and SseEF1A4, represent novel genes that are mainly expressed in gills and skin.The expression profile of the five genes was also studied during larval development,revealing different behaviours.To study the possible regulation of SseEF1A gene expressions by thyroid hormones(THs),larvae were exposed to the goitrogen thiourea (TU).TU-treated larvae exhibited lower SseEF1A4 mRNA levels than untreated controls at both 11 and 15 days after treatment,whereas transcripts of the other four genes remained relatively unchanged. Moreover,addition of exogenous T4 hormone to TU-treated larvae increased significantly the steady- state levels of SseEF1A4 with respect to untreated controls,demonstrating that its expression is up- regulated by THs. Conclusion:We have identified five different eEF1A genes in the Senegalese sole,referred to as SseEF1A1, SseEF1A2,SseEF1A3,SseEF1A4,and Sse42Sp50.The five genes exhibit different expression patterns in tissues and during larval development.TU and T4 treatments demonstrate that SseEF1A4 is up-regulated by THs,suggesting a role in the translational regulation of the factors involved in the dramatic changes that occurs during Senegalese sole metamorphosis.