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Experimental and computational perturbation analysis of enzyme deletions in Drosophila energy metabolism
Author(s) -
Feala Jacob Daniel,
Omens Jeffrey,
Paternostro Giovanni,
McCulloch Andrew
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.756.3
Subject(s) - drosophila melanogaster , hypoxia (environmental) , biology , enzyme , glycolysis , lactate dehydrogenase , gene , energy metabolism , biochemistry , computational biology , biophysics , chemistry , oxygen , organic chemistry , endocrinology
The systems biology approach is complementary to the traditional reductionist approach in that it attempts to use all available data to build increasingly accurate computational models, from which specific new hypotheses can then be generated and tested classically. We use results from various experiments to refine a model of the reaction network of ATP‐generating central metabolism in Drosophila flight muscle. The network, built previously using the annotated genome and the KEGG database of genes and reactions, was made specific for flight muscle by integrating absolute gene expression data for fly thorax, which is composed mostly of flight muscle. Quantitative 1 H spectroscopy and biochemical assays were used to measure accumulation of products, substrate depletion, and ATP concentration over 4 hours of hypoxia (0.5% O 2 ) and anoxia, for wild‐type and several enzyme deletion strains, and the model was fitted to these data by constraining reaction fluxes. Based on production of ATP and protons in the steady‐state computer model, we hypothesized increased hypoxia tolerance in flies deficient in the lactate dehydrogenase (LDH) enzyme. Compared to wild‐type, heterozygous LDH deletions were found to have improved heart rate and faster recovery of mobility after 4 hours hypoxia. The refined version of the model can be used to generate other testable hypothesis regarding chronic hypoxia in Drosophila muscle tissue, or the generic model can be similarly tailored to other specific contexts of interest.