Cloning, structural analysis and mapping of the mouse selenocysteine tRNA([Ser]Sec) gene (Trsp)

Abstract

The tRNA([Ser]Sec) molecule mediates the synthesis of selenoproteins by incorporating selenocysteine into specific UGA codons upon translation of mRNAs that encode selenocysteine-containing proteins. The mouse gene encoding tRNA([Ser]Sec) (Trsp) was isolated from a genomic library and sequenced. The mouse sequence is colinear with its tRNA product, and contains a C to T transition relative to the homologous genes in other vertebrates except rat. Transcriptional control motifs found 5' to the tRNA coding region included a TATA element, a PSE element and an SPH motif which is associated with an octamer motif. A Northern hybridization analysis showed highest expression in the testis, followed by thymus, spleen, kidney, ovary, brain, liver, heart and skeletal muscle. Surprisingly, the expression level was lowest in embryonic stem cells. These results suggest a tissue-specific transcriptional control. Using restriction fragment length variants (RFLVs) in interspecific backcross mice between Mus musculus (C3H strain) and Mus spretus, the Trsp gene was mapped to the proximal region of mouse Chr 7, cosegregating with octamer-binding transcription factor-2 (Otf2).