title: Regulation of NAD synthesis by the trifunctional NadR protein of Salmonella enterica.
abstract: The three activities of NadR were demonstrated in purified protein and assigned to separate domains by missense mutations. The N-terminal domain represses transcription of genes for NAD synthesis and salvage. The C-terminal domain has nicotinamide ribose kinase (NmR-K; EC 2.7.1.22) activity, which is essential for assimilation of NmR, converting it internally to nicotinamide mononucleotide (NMN). The central domain has a weak adenylyltransferase (NMN-AT; EC 2.7.7.1) activity that converts NMN directly to NAD but is physiologically irrelevant. This central domain mediates regulatory effects of NAD on all NadR activities. In the absence of effectors, pure NadR protein binds operator DNA (the default state) and is released by ATP (expected to be present in vivo). NAD allows NadR to bind DNA in the presence of ATP and causes repression in vivo. A superrepressor mutation alters an ATP-binding residue in the central (NMN-AT) domain. This eliminates NMN-AT activity and places the enzyme in its default (DNA binding) state. The mutant protein shows full NmR kinase activity that is 10-fold more sensitive to NAD inhibition than the wild type. It is proposed that NAD and the superrepressor mutation exert their effects by preventing ATP from binding to the central domain.
authors.affiliation: Department of Biology, University of Utah, Salt Lake City, UT 84112, USA.
authors.forename: Julianne H
authors.initials: JH
authors.lastname: Grose
authors.forename: Ulfar
authors.initials: U
authors.lastname: Bergthorsson
authors.forename: John R
authors.initials: JR
authors.lastname: Roth
mesh_terms: D001426:Bacterial Proteins / Q000502:physiology*; D001665:Binding Sites; D004269:DNA, Bacterial / Q000235:genetics; D005809:Genes, Regulator; D009243:NAD / Q000096:biosynthesis*; D009876:Operon; D012097:Repressor Proteins / Q000502:physiology*; D019779:Salmonella enterica / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C117105:NadR protein, bacteria; D012097:Repressor Proteins; D009243:NAD