Summary
A series of Mud1 and Tn10 insertions were identified in the pncA chromosome region of Salmonella typhimurium which is responsible for the production of nicotinamide deamidase. Both pncA (resulting in no nicotinamide deamidase activity) and pncX (resulting in lowered nicotinamide deamidase activity) insertions were constructed. In addition, mutants which could utilize nicotinamide as a sole source of nitrogen were isolated. These mutants, designated pncH, hyperproduce nicotinamide deamidase. Genetic studies utilizing pncX-lacZ and pncA-lacZ operon fusions indicate that pncX::Tn10 insertions reduce transcription of pncA-lac while pncH mutations increase the expression of both pncA-lacZ and pncX-lacZ. The gene order was determined as purB-pncA-pncX-gdh with transcription of both pncA and pncX occurring in the counterclockwise direction. Merodiploid studies suggest a model whereby pncX and pncA form an operon with the major promoter occurring upstream from pncX. A second, weaker promoter for pncA must be situated between pncX and pncA. The pncH mutations appear to occur in the pncX promoter (pncXp) increasing promoter activity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bachmann BJ (1983) Linkage of Escherichia coli K-12, Edition 7. Microbiol Rev 47:180–230
Bochner BR, Huang H, Schieven GL, Ames BN (1980) Positive selection for loss of tetracycline resistance. J Bacteriol 143:926–933
Chan RK, Botstein D, Watanabe T, Ogata Y (1972) Specialized transduction of tetracycline resistance by phage P22 in Salmonella typhimurium. II. Properties of a high transducing lysate. Virology 50:883–898
Chumley FG, Menzel R, Roth JR (1978) Hfr formation directed by Tn10. Genetics 91:639–655
Foster JW, Moat AG (1980) Nicotinamide adenine dinucleotide biosynthesis and pyridine nucleotide cycle metabolism in microbial systems. Microbiol Rev 44:83–105
Foster JW, Kinney DM, Moat AG (1979a) Pyridine nucleotide cycle of Salmonella typhimurium: Isolation and characterization of pncA, pncB and pncC mutants and utilization of exogenous nicotinamide adenine dinucleotide. J Bacteriol 137:1165–1175
Foster JW, Kinney DM, Moat AG (1979b) Pyridine nucleotide cycle of Salmonella typhimurium: Regulation of nicotinic acid phosphoribosyltransferace and nicotinamide deamidase. J Bacteriol 138:957–961
Fukada M (1983) Superhelical model of DNA for regulation of gene expression. J Theor Biol 100:239–254
Holley EA, Foster JW (1982) Bacteriophage P22 as a vector for Mu mutagenesis in Salmonella typhimurium: Isolation of nalac and pnc-lac gene fusions. J Bacteriol 152:959–962
Hong JS, Ames BN (1971) Localized mutagenesis of any specific small regions of the bacterial chromosome. Proc Natl Acad Sci USA 68:3158–3162
Hughes KT, Roth JR (1984) Conditionally, transposition defective derivative of Mud1 (Amp Lac). J Bacteriol 159:130–137
Hughes KT, Cookson BT, Ladika D, Olivera BM, Roth JR (1983) 6-Aminoicotinamide-resistant mutants of Salmonella typhimurium. J Bacteriol 154:1126–1136
Kinney DM, Foster JW (1985) Identification of a cis-acting regulatory region in the pncB locus of Salmonella typhimurium. Mol Gen Genet 199:512–517
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Miller ES, Brenchley JE (1984) Cloning and characterization of gdh, the structural gene for glutamate dehydrogenase of Salmonella tyhpimurium. J Bacteriol 157:171–178
Miller JH (ed) (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Neidhardt FC, Bloch PL, Smith DF (1974) Culture medium for enterobacteria. J Bacteriol 119:736–747
Pardee AB, Benz EJ, ST Peter DA, Krieger JN, Neuth M, Trieshman HW (1971) Hyperproduction and purification of nicotinamide deamidase a microconstitutive enzyme of Escherichia coli. Biol Chem 246:6792–6796
Riley M, Anilionis A (1978) Evolution of the bacterial genome. Ann Rev Microbiol 32:519–560
Rosenfeld SA, Dendinger SM, Murphy CH, Brenchley JE (1982) Genetic characterization of the glutamate dehydrogenase gene (gdh) of Salmonella typhimurium. J Bacteriol 150:795–803
Sanderson KE, Roth JR (1983) Linkage map of Salmonella typhimurium, Edition VI. Microbiol Rev 47:410–453
Smith JR (1981) DNA supercoiling: another level for regulating gene expression. Cell 24:599–600
Sternglanz RD, Nardo S, Voelkel KA, Nishimura Y, Hirota Y, Becherer K, Zumstein L, Wang JC (1981) Mutations in the gene coding for Escherichia coli DNA topoisomerase I affecting transcription and transposition. Proc Natl Acad Sci USA 78:2747–2751
Tirgari S, Spector MP, Foster JW (1986) Genetics of NAD metabolism in Salmonella typhimurium and cloning of the nadA and pnuC loci. J Bacteriol 167:1086–1088
Vogel HJ, Bonner DM (1956) Acetyl ornithinase of Escherichia coli: Partial purification and some properties. J Biol Chem 93:237–244
Author information
Authors and Affiliations
Additional information
Communicated by J. Lengeler
Rights and permissions
About this article
Cite this article
Hill-Chappell, J.M., Spector, M.P. & Foster, J.W. The pyridine nucleotide cycle of Salmonella typhimurium: Genetic characterization of the pncXA operon. Mol Gen Genet 205, 507–514 (1986). https://doi.org/10.1007/BF00338090
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00338090