Proteomic analysis uncovers a metabolic phenotype in C. elegans after nhr-40 reduction of function |
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Authors: | Pohludka Michal Simeckova Katerina Vohanka Jaroslav Yilma Petr Novak Petr Krause Michael W Kostrouchova Marta Kostrouch Zdenek |
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Institution: | a Charles University in Prague, 1st Faculty of Medicine, Institute of Inherited Metabolic Disorders, Laboratory of Molecular Pathology, Ke Karlovu 2, CZ-128 01 Prague, Czech Republic b Charles University in Prague, 1st Faculty of Medicine, Institute of Inherited Metabolic Disorders, Laboratory of Molecular Biology and Genetics, Ke Karlovu 2, CZ-128 01 Prague, Czech Republic c Laboratory of Molecular Structure Characterization, Institute of Microbiology, Czech Academy of Sciences, Videnska 1083, CZ-142 20 Prague, Czech Republic d Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892, USA |
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Abstract: | Caenorhabditis elegans has an unexpectedly large number (284) of genes encoding nuclear hormone receptors, most of which are nematode-specific and are of unknown function. We have exploited comparative two-dimensional chromatography of synchronized cultures of wild type C. elegans larvae and a mutant in nhr-40 to determine if proteomic approaches will provide additional insight into gene function. Chromatofocusing, followed by reversed-phase chromatography and mass spectrometry, identified altered chromatographic patterns for a set of proteins, many of which function in muscle and metabolism. Prompted by the proteomic analysis, we find that the penetrance of the developmental phenotypes in the mutant is enhanced at low temperatures and by food restriction. The combination of our phenotypic and proteomic analysis strongly suggests that NHR-40 provides a link between metabolism and muscle development. Our results highlight the utility of comparative two-dimensional chromatography to provide a relatively rapid method to gain insight into gene function. |
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