Superfund Research Program
Effects Related Biomarkers of Toxic Exposures
Project Leader: Terrance J. Kavanagh
Grant Number: P42ES004696
Funding Period: 1995 - 2006
As part of efforts to characterize regulatory regions of mGLCLC and mGLCLR, project investigators have subcloned the 5' regions of mouse GLCLR and GLCLC into reporter vectors (including luciferase and green fluorescent protein reporters) to assess in mammalian cell culture experiments, and to clarify their role in regulating basal and inducible mRNA expression. Evidence to date indicates that the mouse GLCLC 5' promoter is a strong constitutive promoter in Hepa-1 mouse liver cells, but that regions 5' of the ARE cooperate in inducibility.
Further work is focusing on post-translational modifications of GLCL resulting from oxidative treatments, such as methyl mercury (MeHg). Using Northern analysis, in situ hybridization, Western analysis, immunocytochemistry and HPLC assays, investigators are examining the effects of MeHg exposure on GLCL expression and activity and GSH levels in various tissues in mice, to localize expression changes at the individual tissue/cell level. Preliminary findings suggest that GLCL may be down-regulated in certain tissues (lung, intestine) with MeHg exposure as opposed to being up-regulated in kidney, liver and brain. Other directions that are being pursued include investigation of the role of the GAG repeat polymorphism in the 5' region of the human GLCL gene in idiopathic pulmonary fibrosis, a disease considered likely to have a strong oxidative stress pathogenic component.