| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Submitted on April 22, 2004
R & D, Xantos Biomedicine AG, Munich 81377
Corresponding Author: u.brinkmann{at}xantos.de
We describe a novel genetic screen that is performed by transfecting every individual clone of an expression clone collection into a separate population of cells in a high throughput mode. We combined high throughput functional genomics with experimental validation to discover human genes which ameliorate cytotoxic responses of neuronal HT-22 cells upon exposure to oxidative stress. A collection of 5000 human cDNAs in mammalian expression vectors were individually transfected into HT-22 cells, which were then exposed to H2O2. Five genes were found that are known to be involved in pathways of detoxification of peroxide (catalase, glutathione peroxidase-1, peroxiredoxin-1, peroxiredoxin-5 and nuclear factor erythroid-derived 2-like 2). The presence of those genes in our ‘hit list’ validates our screening platform. In addition, a set of candidate genes were found that have not been previously described to be involved in detoxification of peroxide. One of these genes which was consistently found to reduce H2O2 - induced toxicity in HT-22, was GFPT2. This gene is expressed at significant levels in the central nervous system and encodes glutamine-fructose-6-phosphate transaminase 2, a rate limiting enzyme in hexosamine biosynthesis. GFPT has recently also been shown to ameliorate the toxicity of methylmercury in Saccharomyces cerevisiae. Methylmercury causes neuronal cell death in part by protein modification as well as enhancing the production of ROS. The protective effect of GFPT2 against H2O2-toxicity in neuronal HT-22 cells may be similar to its protection against methylmercury in yeast. Thus, GFPT appears to be conserved among yeast and men as a critical target of methylmercury and ROS induced cytotoxicity.
Revised on June 3, 2004
Accepted on June 4, 2004
High throughput functional genomics identifies genes which ameliorate toxicity due to oxidative stress in neuronal HT22 cells: GFPT2 protects cells against peroxide
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  R. C. Cumming, R. Dargusch, W. H. Fischer, and D. Schubert Increase in Expression Levels and Resistance to Sulfhydryl Oxidation of Peroxiredoxin Isoforms in Amyloid beta-Resistant Nerve Cells J. Biol. Chem., October 19, 2007; 282(42): 30523 - 30534. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Liu, J. T. Kern, J. R. Walker, J. A. Johnson, P. G. Schultz, and H. Luesch A genomic screen for activators of the antioxidant response element PNAS, March 20, 2007; 104(12): 5205 - 5210. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Korherr, H. Gille, R. Schafer, K. Koenig-Hoffmann, J. Dixelius, K. A. Egland, I. Pastan, and U. Brinkmann Identification of proangiogenic genes and pathways by high-throughput functional genomics: TBK1 and the IRF3 pathway. PNAS, March 14, 2006; 103(11): 4240 - 4245. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. C. Cooksey, S. Pusuluri, M. Hazel, and D. A. McClain Hexosamines regulate sensitivity of glucose-stimulated insulin secretion in {beta}-cells Am J Physiol Endocrinol Metab, February 1, 2006; 290(2): E334 - E340. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Korn, S. Rohrig, S. Schulze-Kremer, and U. Brinkmann Common denominator procedure: a novel approach to gene-expression data mining for identification of phenotype-specific genes Bioinformatics, June 1, 2005; 21(11): 2766 - 2772. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Journal of Biological Chemistry |
| Journal of Lipid Research | ASBMB Today |
