Nuclear functions in cells depend upon critical thiols that are subject to oxidative damage by environmental, occupational and therapeutic chemicals. Thioredoxin-1 (Trx1) and glutathione (GSH) are biological thiols that protect against oxidants. These thiols also function in redox signaling and control. The purpose of this proposal is to gain an understanding of the distinct functions of Trx1 and GSH in protection against oxidative stress in cell nuclei.
Aim 1 is focused on the quantitative importance of the nuclear Trx1 system compared to cytoplasmic Trx1 and GSH-dependent systems for protection against oxidative stress. Cell models will be used to examine nuclear responses to exogenous H2O2, endogenously nuclear-generated H2O2 and oxidant chemicals. Novel redox western blot and mass spectrometry proteomic techniques will be used to determine the redox state of nuclear Trx1 and S-glutathionylation of nuclear proteins. Nuclear-targeted Trx1 (NLS-Trx1) and dominant negative forms will be used to control the nuclear Trx1 system;NLS-peroxiredoxin-1 (NLS-Prx1) will be used to enhance nuclear peroxide metabolism;and cellular GSH will be modified by control of synthesis and by selective depletion. Nuclear-targeted (NLS-) D- amino acid oxidase with added D-amino acids will be used to stimulate intranuclear H2O2 generation. Toxicants will be studied to test for selective nuclear resistance/sensitivity to oxidative stress.
Specific Aim 2 is to develop transgenic mouse models expressing fusion proteins with nuclear localization and nuclear export signals (NES) fused to human Trx1 (NLS-hTrx1 and NES-hTrx1) and use these for in vivo toxicologic studies of nuclear oxidative stress. Transgenic mice will be characterized for nuclear and cytoplasmic redox under control conditions and with an established in vivo toxicologic challenge with the cardiotoxicant, adriamycin. The results of these studies will provide the first test of the hypothesis that enhanced nuclear antioxidant systems protect against toxicity in vivo.
Specific Aim 3 is to use mass spectrometry-based redox proteomic techniques to identify nuclear proteins controlled by the Trx1 and GSH systems and to identify nuclear proteins that are selectively vulnerable to oxidative stress. The successful completion of these aims will provide fundamental new knowledge concerning the quantitative importance of major thiol/disulfide control systems in nuclei in protection against oxidative stress from environmental and therapeutic toxicities.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES011195-10
Application #
8055488
Study Section
Xenobiotic and Nutrient Disposition and Action Study Section (XNDA)
Program Officer
Carlin, Danielle J
Project Start
2001-09-01
Project End
2012-03-31
Budget Start
2011-04-01
Budget End
2012-03-31
Support Year
10
Fiscal Year
2011
Total Cost
$330,651
Indirect Cost
Name
Emory University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
066469933
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Soltow, Quinlyn A; Strobel, Frederick H; Mansfield, Keith G et al. (2013) High-performance metabolic profiling with dual chromatography-Fourier-transform mass spectrometry (DC-FTMS) for study of the exposome. Metabolomics 9:S132-S143
Uppal, Karan; Soltow, Quinlyn A; Strobel, Frederick H et al. (2013) xMSanalyzer: automated pipeline for improved feature detection and downstream analysis of large-scale, non-targeted metabolomics data. BMC Bioinformatics 14:15
Go, Young-Mi; Orr, Michael; Jones, Dean P (2013) Increased nuclear thioredoxin-1 potentiates cadmium-induced cytotoxicity. Toxicol Sci 131:84-94
Go, Young-Mi; Jones, Dean P (2013) Thiol/disulfide redox states in signaling and sensing. Crit Rev Biochem Mol Biol 48:173-81
Jones, Dean P; Park, Youngja; Ziegler, Thomas R (2012) Nutritional metabolomics: progress in addressing complexity in diet and health. Annu Rev Nutr 32:183-202
Roede, James R; Go, Young-Mi; Jones, Dean P (2012) Redox equivalents and mitochondrial bioenergetics. Methods Mol Biol 810:249-80
Jin, Ran; Le, Ngoc-Anh; Liu, Shuling et al. (2012) Children with NAFLD are more sensitive to the adverse metabolic effects of fructose beverages than children without NAFLD. J Clin Endocrinol Metab 97:E1088-98
Liang, Yongliang; Roede, James R; Dikalov, Sergey et al. (2012) Determination of ebselen-sensitive reactive oxygen metabolites (ebROM) in human serum based upon N,N'-diethyl-1,4-phenylenediamine oxidation. Clin Chim Acta 414:1-6
Park, Youngja H; Lee, Kichun; Soltow, Quinlyn A et al. (2012) High-performance metabolic profiling of plasma from seven mammalian species for simultaneous environmental chemical surveillance and bioeffect monitoring. Toxicology 295:47-55
Jones, Dean P; Go, Young-Mi (2011) Mapping the cysteine proteome: analysis of redox-sensing thiols. Curr Opin Chem Biol 15:103-12

Showing the most recent 10 out of 66 publications