Cloning and expression of a cDNA for a mammalian type III iodothyronine deiodinase

The type III iodothyronine deiodinase metabolizes the active thyroid hormones thyroxine and 3,5,3'-triiodothyronine to inactive compounds. Recently, we have characterized a Xenopus laevis cDNA (XL-15) that encodes a selenoprotein with type III deiodinase activity (St. Germain, D.L., Schwartzman, R., Croteau, W., Kanamori, A., Wang, Z., Brown, D.D., and Galton, V.A. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 7767-7771). Using the XL-15 as a probe, we screened a rat neonatal skin cDNA library. Among the clones isolated was one (rNS43-1) which contained a 2.1-kilobase pair cDNA insert that manifested significant homology to both the XL-15 and the G21 rat type I deiodinase cDNAs, including the presence of an in-frame TGA codon. Expression studies demonstrated that the rNS43-1 cDNA encodes a protein with 5-, but not 5'-, deiodinase activity that is resistant to inhibition by propylthiouracil and aurothioglucose. Northern analysis demonstrated a pattern of tissue expression in the rat consistent with that of the type III deiodinase and site directed mutagenesis confirmed that the TGA triplet codes for selenocysteine. We conclude that the rNS43-1 cDNA encodes the rat type III deiodinase and that the types I and III deiodinases present in amphibians and mammals constitute a family of conserved selenoproteins important in the metabolism of thyroid hormones.