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J Biol Chem, Vol. 274, Issue 26, 18729-18734, June 25, 1999

Selenium Metabolism in Drosophila
CHARACTERIZATION OF THE SELENOCYSTEINE tRNA POPULATION

Xuan Zhou, Sang Ick Park^§, Mohamed E. Moustafa, Bradley A. Carlson, Pamela F. Crain^¶, Alan M. Diamond, Dolph L. Hatfield, and Byeong Jae Lee^§

From the  Section on the Molecular Biology of Selenium, Laboratory of Basic Research, NCI, National Institutes of Health, Bethesda, Maryland 20892, the ^¶ Department of Medicinal Chemistry, University of Utah, Salt Lake City, Utah 84112, the ^§ Laboratory of Molecular Genetics, Institute for Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Korea, and the  Department of Human Nutrition and Dietetics, University of Illinois at Chicago, Chicago, Illinois 60612

The selenocysteine (Sec) tRNA population in Drosophila melanogaster is aminoacylated with serine, forms selenocysteyl-tRNA, and decodes UGA. The K_m of Sec tRNA and serine tRNA for seryl-tRNA synthetase is 6.67 and 9.45 nM, respectively. Two major bands of Sec tRNA were resolved by gel electrophoresis. Both tRNAs were sequenced, and their primary structures were indistinguishable and colinear with that of the corresponding single copy gene. They are 90 nucleotides in length and contain three modified nucleosides, 5-methylcarboxymethyluridine, N⁶-isopentenyladenosine, and pseudouridine, at positions 34, 37, and 55, respectively. Neither form contains 1-methyladenosine at position 58 or 5-methylcarboxymethyl-2'-O-methyluridine, which are characteristically found in Sec tRNA of higher animals. We conclude that the primary structures of the two bands of Sec tRNA resolved by electrophoresis are indistinguishable by the techniques employed and that Sec tRNAs in Drosophila may exist in different conformational forms. The Sec tRNA gene maps to a single locus on chromosome 2 at position 47E or F. To our knowledge, Drosophila is the lowest eukaryote in which the Sec tRNA population has been characterized to date.

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