The objective of these studies will be to determine how the levels of aminoacyl-tRNA synthetases are regulated in a higher eukaryote, Bombyx mori. This organism will be employed because its silkgland specializes its protein synthetic apparatus for the synthesis of two unusual protein (fibroin and sericin). The goal of the project will be accomplished by purifying three aminoacyl-tRNA synthetases (for glycine, alanine, and serine) and using the purified proteins to develop quantitative immunological assays for the enzyme protein. The levels of these enzymes, will be determined in the silkgland and other organs to determine if the observed specialization of tRNAs for fibroin and sericin production is reflected in the levels of the cognate aminoacyl-tRNA synthetases. This approach will make it possible to determine if the observed changes in enzyme activity during silkgland development result from changes in enzyme specific activity or from increased levels of enzyme protein. The long term goal of these studies will be the elucidation of basic cellular processes that control the levels of these enzymes. An additional objective will be the cloning of aminoacyl-tRNA synthetase genes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032410-08
Application #
3281205
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1988-09-01
Project End
1991-06-30
Budget Start
1989-07-01
Budget End
1990-06-30
Support Year
8
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Toledo
Department
Type
Schools of Medicine
DUNS #
807418939
City
Toledo
State
OH
Country
United States
Zip Code
43614
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Allen, K N; Lavie, A; Glasfeld, A et al. (1994) Role of the divalent metal ion in sugar binding, ring opening, and isomerization by D-xylose isomerase: replacement of a catalytic metal by an amino acid. Biochemistry 33:1488-94
Lavie, A; Allen, K N; Petsko, G A et al. (1994) X-ray crystallographic structures of D-xylose isomerase-substrate complexes position the substrate and provide evidence for metal movement during catalysis. Biochemistry 33:5469-80
Allen, K N; Lavie, A; Farber, G K et al. (1994) Isotopic exchange plus substrate and inhibition kinetics of D-xylose isomerase do not support a proton-transfer mechanism. Biochemistry 33:1481-7
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Chang, P K; Dignam, J D (1990) Primary structure of alanyl-tRNA synthetase and the regulation of its mRNA levels in Bombyx mori. J Biol Chem 265:20898-906
Viswanathan, S; Dignam, J D (1988) Seryl-tRNA synthetase from Bombyx mori. Purification and properties. J Biol Chem 263:535-41
Viswanathan, S; Dignam, S S; Dignam, J D (1988) Control of the levels of alanyl-, glycyl-, and seryl-tRNA synthetases in the silkgland of Bombyx mori. Dev Biol 129:350-7