The long-term objective of our research is to understand the molecular mechanisms of cell death as a consequence of harmful environmental conditions. There appear to be several distinct circumstances under which cell death takes place in multicellular organisms. For example, apoptosis (programmed cell death) is known to play an important role in embryonic development, normal cell turnover, and hormone-dependent atrophy. Environmentally adverse chemicals can lead to cytotoxicity and, ultimately, cell death. Although apoptosis and cytotoxicity appear to occur for many reasons and respond to a variety of signals, it has become increasingly evident that there exist common steps in the pathways leading to cell death. Chemicals causing oxidative damage are metabolized by """"""""Phase I"""""""" (oxygenation) and """"""""Phase II"""""""" (conjugate) enzymes. We propose to use the aromatic hydrocarbon-responsive [Ah] gene battery as a model system for studying the cellular response of protection against oxidative stress and cell death. In the mouse, the [Ah] battery comprises at least six genes: two Phase I genes, Cypla1 and Cypla2; and four Phase II genes, Nmo-1, Aldh- 1, Ugt-1 and Gt-1. The radiation-induced deletion homozygote c14CoS/c14CoS mouse (albino phenotype) is missing about 1 centiMorgan of chromosome 7, dies during the first 18 h post partum, and exhibits marked activation of the Nmo-1 gene, increases in Aldh-1 mRNA, and elevated Ugt-1 and Gt-1 enzyme activities. Interestingly, in the 14CoS/14CoS fetus and newborn, three growth arrest- and DNA damage-inducible (gadd) genes are also activated. We postulate that the region of mouse chromosome 7 missing in the 14CoS/14CoS mouse contains a """"""""master switch"""""""" gene, which we have designated Nmo-1n, encoding a trans-acting factors(s) that is a negative effector of the Nmo-1 gene. Under normal conditions, Nmo-1n suppresses an unknown number of genes. In response to environmental adversity such as oxidative stress, this gene releases its negative control on the [Ah] battery Phase II genes, and perhaps the gadd genes, thereby allowing all of these genes it become expressed. This response is independent of Phase I (Cypla1 and Cypla2) gene expression. Whether this region on chromosome 7 represents a single gene, Nmo-1a, controlling both the [Ah] battery Phase II genes and the gadd genes, or whether there are two or more gene controlling the [Ah] battery Phase II genes and the gadd genes, will require further work. Our first goal is to clone, sequence and characterize by functional analysis the murine Nmo-1 and Aldh-1 genes and regulatory regions. We then propose to use this information for cloning and characterizing the murine Nmo-1n gene. Knowledge about the Nmo-1n gene and its product will surely help clarify the functional relevance of common pathways involved in protection against oxidative damage and cell death.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG009235-03
Application #
3121034
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1990-09-01
Project End
1995-08-31
Budget Start
1992-09-01
Budget End
1993-08-31
Support Year
3
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Cincinnati
Department
Type
Schools of Medicine
DUNS #
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Dalton, Timothy P; Chen, Ying; Schneider, Scott N et al. (2004) Genetically altered mice to evaluate glutathione homeostasis in health and disease. Free Radic Biol Med 37:1511-26
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Yang, Yi; Dieter, Matthew Z; Chen, Ying et al. (2002) Initial characterization of the glutamate-cysteine ligase modifier subunit Gclm(-/-) knockout mouse. Novel model system for a severely compromised oxidative stress response. J Biol Chem 277:49446-52
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Nebert, D W; Roe, A L; Dieter, M Z et al. (2000) Role of the aromatic hydrocarbon receptor and [Ah] gene battery in the oxidative stress response, cell cycle control, and apoptosis. Biochem Pharmacol 59:65-85
Dalton, T P; Solis, W A; Nebert, D W et al. (2000) Characterization of the MTF-1 transcription factor from zebrafish and trout cells. Comp Biochem Physiol B Biochem Mol Biol 126:325-35
Dalton, T P; Dieter, M Z; Yang, Y et al. (2000) Knockout of the mouse glutamate cysteine ligase catalytic subunit (Gclc) gene: embryonic lethal when homozygous, and proposed model for moderate glutathione deficiency when heterozygous. Biochem Biophys Res Commun 279:324-9
Vasiliou, V; Buetler, T; Eaton, D L et al. (2000) Comparison of oxidative stress response parameters in newborn mouse liver versus simian virus 40 (SV40)-transformed hepatocyte cell lines. Biochem Pharmacol 59:703-12

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