Research
Interests
I have been interested in the regulation of transcription by eukaryotic RNA
polymerase III. The genes encoding alanine tRNA in the silkworm, Bombyx mori are a striking example of regulated polymerase III activity. One class (C) is
transcribed constitutively; the other (SG) is transcribed only in cells of the
silk gland, where it contributes to efficient silk production. Our goals were
to understand how differences in affinity for particular polypeptides and the
spatial arrangement of bound proteins in the upstream promoter element resulted
in the differential transcription of B. mori's two kinds of tRNA Ala
genes.
Our approach included analyzing the function of cloned genes by transcribing
them in vitro then introducing them into insect cells and site-directed mutagenesis
in which we identified sequence and structural differences in the SG promoter.
Publications
Ouyang, C., J. Martinez, L.S. Young, and K.U. Sprague. (2000) TBP-TATA interaction
is a key determinant of differential transcription of silkworm constitutive
and silkgland-specific tRNA Ala genes. Mol.
Cell Biol. 20, 1329-43.
Trivedi, A., L.S. Young, C. Ouyang, D.L. Johnson, and K.U. Sprague. (1999)
A TATA element is required for tRNA promoter activity and confers TATA-binding
responsiveness in Drosophila Schneider-2 cells. J.
Biol. Chem. 274, 11369-75.
Ouyang, C. and K.U. Sprague.(1998) Cloning and characterization of the TATA-binding
protein of the silkworm Bombyx mori. Gene 221, 207-13.
Young, L.S., N. Ahnert, and K.U. Sprague. (1996) Silkworm TFIIIB binds both
constitutive and silkgland-specific tRNAAla genes, but protects only the constitutive
promoter from DNase I cleavage. Mol. Cell. Biol. 16, 1256-66.
Smith, T.P., L.S. Young, and K.U. Sprague. (1995) Silkworm TFIIIA requires
additional class III factors for commitment to transcription complex assembly
on a 5S RNA gene, Nucl. Acids Res. 23, 1244-51.
Sprague, K.U. (1995) Transcription of eukaryotic tRNA genes. In: Transfer
RNA, D. Söll and U.L. RajBhandary (eds.), ASM Press, Chapter
4, pp 31-50.
Sprague, K.U. (1995) Control of transcription by Bombyx mori RNA polymerase
III. In: Molecular Model Systems in the Lepidoptera, M. Goldsmith and
A. Wilkins (eds.), Cambridge University Press, Chapter 19, pp 273-91.
Dunstan, H.M., L.S. Young, and K.U. Sprague. (1994) tRNA plays an indirect
role in silkworm class III transcription in vitro and inhibits low frequency
DNA cleavage, Mol. Cell. Biol. 14, 3596-603.
Sullivan, H.S., L.S. Young, C.N. White, and K.U. Sprague. (1994) Silkgland-specific
tRNAAla genes interact more weakly than constitutive tRNAAla genes
with silkworm TFIIIB and polymerase III fractions. Mol. Cell. Biol. 14,
1806-14.
Dunstan, H.M., L.S. Young, and K.U. Sprague. (1994) TFIIIR is an isoleucine
tRNA. Mol. Cell. Biol. 14, 3588-95.
Sullivan, H., L.S. Young, C.N. White, and K.U. Sprague. (1994) Silk gland-specific
tRNAAla genes interact more weakly than constitutive tRNAAla genes with silkworm
TFIIIB and polymerase III fractions. Mol. Cell. Biol. 14, 1806-14.
Palida, F.A., C. Hale, and K.U. Sprague. (1993) Transcription of a silkworm
tRNAAlaC gene is directed by two AT-rich upstream sequence elements. Nucl.
Acids Res. 21, 5875-81.
Dieci, G., L. Duimio, F. Coda-Zabetta, K.U. Sprague, and S. Ottonello. (1993)
A novel RNA polymerase III transcription factor fraction that is not required
for template commitment. J. Biol. Chem. 268, 11199-207.
Young, L.S., D.H. Rivier, and K.U. Sprague. (1991) Sequences far downstream
from the classical tRNA. Promoter elements bind RNA polymerase III transcription
factors. Mol. Cell. Biol. 11, 1382-92.
Young, L.S., N. Takahashi, and K.U. Sprague. (1986) Upstream sequences
confer distinctive transcriptional properties on genes encoding silkgland-specific
tRNAAla. Proc. Natl. Acad. Sci. USA 83, 374-8. |
