Abstract
Selenocysteine (Sec) tRNA has been described as the key molecule and the central component in selenoprotein (SP) biosynthesis and is the only known tRNA that governs the expression of an entire class of proteins, the SPs. This provides a unique opportunity to study the expression of SPs by manipulating the levels and characteristics of Sec tRNA. A cDNA encoding a protein that binds to the activator element of Sec tRNA was cloned from X. laevis and named Staf (for Sec tRNA gene transcription activating factor) (Adachi, et al. J Biol Chem 273:, 1998). Herein, we generated a transgenic mouse harboring multiple copies of a transgene carrying a 14bp deletion in the Staf binding region of the Sec tRNA gene (trsp), which is located approximately −200bp upstream of trsp. Transgenic mice were bred with a trsp knockout mouse to remove endogenous wild type trsp. Mice dependent on the mutant transgene showed ~80% decrease in the levels of Sec tRNA in liver. Se75-labeling and western blot analysis revealed that several SPs were affected by this mutation to varying degrees depending on tissue type. The mutant mice showed decreased size and body weight and had a lifespan of less than 30 days. Thus, the Staf binding region of trsp is essential for proper amounts of Sec tRNA transcription. These and other recent findings will be presented. This research was supported by the Intramural Research Program of the NIH, NCI, CCR.
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