title: Computational prediction and experimental verification of the gene encoding the NAD+/NADP+-dependent succinate semialdehyde dehydrogenase in Escherichia coli.
abstract: Although NAD(+)-dependent succinate semialdehyde dehydrogenase activity was first described in Escherichia coli more than 25 years ago, the responsible gene has remained elusive so far. As an experimental proof of concept for a gap-filling algorithm for metabolic networks developed earlier, we demonstrate here that the E. coli gene yneI is responsible for this activity. Our biochemical results demonstrate that the yneI-encoded succinate semialdehyde dehydrogenase can use either NAD(+) or NADP(+) to oxidize succinate semialdehyde to succinate. The gene is induced by succinate semialdehyde, and expression data indicate that yneI plays a unique physiological role in the general nitrogen metabolism of E. coli. In particular, we demonstrate using mutant growth experiments that the yneI gene has an important, but not essential, role during growth on arginine and probably has an essential function during growth on putrescine as the nitrogen source. The NADP(+)-dependent succinate semialdehyde dehydrogenase activity encoded by the functional homolog gabD appears to be important for nitrogen metabolism under N limitation conditions. The yneI-encoded activity, in contrast, functions primarily as a valve to prevent toxic accumulation of succinate semialdehyde. Analysis of available genome sequences demonstrated that orthologs of both yneI and gabD are broadly distributed across phylogenetic space.
authors.affiliation: Institute of Molecular Systems Biology, ETH Zurich, CH-8093 Zurich, Switzerland.
authors.forename: Tobias
authors.initials: T
authors.lastname: Fuhrer
authors.forename: Lifeng
authors.initials: L
authors.lastname: Chen
authors.forename: Uwe
authors.initials: U
authors.lastname: Sauer
authors.forename: Dennis
authors.initials: D
authors.lastname: Vitkup
mesh_terms: D003198:Computer Simulation; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D050644:Succinate-Semialdehyde Dehydrogenase / Q000235:genetics* / Q000378:metabolism
chemical_list: D050644:Succinate-Semialdehyde Dehydrogenase