Molecular cloning of vitellogenin gene promoters and in vitro and in vivo transcription profiles following estradiol-17β administration in the cutthroat trout

Abstract

Transcription of vitellogenin (vtg) genes are initiated when estradiol-17β (E₂)-estrogen receptor (ER) complexes bind estrogen response elements (ERE) located in the gene promoter region. Transcriptional regulation of dual vtg subtypes (major salmonid A-type vtg: vtgAs; minor C-type vtg: vtgC) by E₂ was investigated under co-expression of a potential major transcriptional factor, erα1, in cutthroat trout. Two forms of trout vtgAs promoters (1 and 2) and one vtgC promoter were sequenced. These promoters structurally differ based on the number of EREs present. The vtgAs promoter 1 exhibited the highest maximal transcriptional activity by in vitro gene reporter assays. The concentration of E₂ that induces 50% of gene reporter activity (half-maximal effective concentrations, EC₅₀) was similar among all vtg promoters and also to the EC₅₀ of E₂ administered to induce vtg transcription in vivo. This study revealed a difference in transcriptional properties of multiple vtg promoters for the first time in a salmonid species, providing the basis to understand mechanisms underlying regulation of vitellogenesis via dual vtg gene expression.

Keywords: Estrogen receptor; Estrogen response element; Gene promoter; Reporter gene assay; Vitellogenin.