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J Biol Chem, Vol. 274, Issue 16, 11369-11375, April 16, 1999

A TATA Element Is Required for tRNA Promoter Activity and Confers TATA-binding Protein Responsiveness in Drosophila Schneider-2 Cells

Alpa Trivedi, Lisa S. Young^§, Ching Ouyang^¶, Deborah L. Johnson, and Karen U. Sprague^§

From the  Departments of Molecular Pharmacology and Biochemistry, Schools of Pharmacy and Medicine, University of Southern California, Los Angeles, California 90033 and the ^§ Institute of Molecular Biology, the  Department of Biology, and the ^¶ Department of Physics, University of Oregon, Eugene, Oregon 97403

In contrast to yeast and mammalian systems, which depend principally on internal promoter elements for tRNA gene transcription, insect systems require additional upstream sequences. To understand the function of the upstream sequences, we have asked whether the Bombyx mori tRNA_C^Ala and tRNA_SG^Ala genes, which are absolutely dependent on these sequences in vitro, also require them for transcription in vivo. We introduced wild-type and mutant versions of the Bombyx tRNA^Ala genes into Drosophila Schneider-2 cells and found that the tRNA_C^Ala gene is efficiently transcribed and that its transcription depends strongly on the distal segment of its upstream promoter. In contrast, the tRNA_SG^Ala gene is inefficiently transcribed, and this inefficiency results from lack of a specific sequence within the distal tRNA_C^Ala upstream promoter. This sequence, 5'-TTTATAT-3', is sufficient to increase the activity of the tRNA_SG^Ala promoter to that of the tRNA_C^Ala promoter. Moreover, promoters containing the 5'-TTTATAT-3' element are stimulated by increased levels of cellular TATA-binding protein. Together these results indicate that, in insect cells, a TATA-like element is specifically required to form functional TATA-binding protein-containing complexes that promote efficient transcription of tRNA genes.

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