Student Research: Juliane P. Hill

, , 1993
Faculty Advisor: Terrance J. Kavanagh

Cloning and Partial Characterization of the Mouse Gamma-Glutamylcysteine Synthetase Gene


Gamma-glutamylcysteine (g-GCS) synthetase is the rate-limiting enzyme in a two step pathway that synthesizes glutathione (GSH). Isolated from rate kidney, the g-GCS enzyme consists of two subunits which can be dissociated under denaturing or nondenaturing conditions. The heavy subunit displays all the catalytic activity of the isolated enzyme, as well as feedback inhibition by GSH. The light subunit is enzymatically inactive and its presence does not alter the activity or stability of the heavy subunit.

The overiding hypothesis for this project is that: The gamma-glutamylcysteine synthetase (g-GCS) gene promoter region containes nucleotide sequences which confer inducibility for g-GCS messenger RNA when cells are subject to oxidative stress or chemicals which deplete glutathione (GSH).

The specific objective of this thesis project are to: 1) clone and begin characterization of the moouse genomic g-GCS gene, and 2) develop a probe to be used from chromosome mapping of the g-GCS gene. The training goal of this project was to learn some basic molecular biology techniques and apply them to toxicological research.

Taken from the beginning of thesis.