Genetics, Vol 143, 37-44, Copyright © 1996

INVESTIGATIONS

Genetic Analysis of Metabolic Crosstalk and Its Impact on Thiamine Synthesis in Salmonella typhimurium

L. Petersen, J. Enos-Berlage and D. M. Downs
Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706

The first five steps in de novo purine biosynthesis are involved in the formation of the 4-amino-5-hydroxymethyl-2-methyl pyrimidine (HMP) moiety of thiamine. We show here that the first enzyme in de novo purine biosynthesis, PurF, is required for thiamine synthesis during aerobic growth on some but not other carbon sources. We show that PurF-independent thiamine synthesis depends on the recently described alternative pyrimidine biosynthetic (APB) pathway. Null mutations in zwf (encoding glucose-6-P dehydogenase), gnd (encoding gluconate-6-P dehydrogenase), purE (encoding aminoimidazole ribo-nucleotide carboxylase), and purR (encoding a regulator of gene expression) were found to affect the function of the APB pathway. A model is presented to account for the involvement of these gene products in thiamine biosynthesis via the APB pathway. Results presented herein demonstrate that function of the APB pathway can be prevented either by blocking intermediate formation or by diverting intermediate(s) from the pathway. Strong genetic evidence supports the conclusion that aminoimidazole ribotide (AIR) is an intermediate in the APB pathway.

This article has been cited by other articles:

				J. M. Boyd, J. A. Lewis, J. C. Escalante-Semerena, and D. M. Downs Salmonella enterica Requires ApbC Function for Growth on Tricarballylate: Evidence of Functional Redundancy between ApbC and IscU J. Bacteriol., July 1, 2008; 190(13): 4596 - 4602. [Abstract] [Full Text] [PDF]

				I. Ramos, E. I. Vivas, and D. M. Downs Mutations in the Tryptophan Operon Allow PurF-Independent Thiamine Synthesis by Altering Flux In Vivo J. Bacteriol., February 1, 2008; 190(3): 815 - 822. [Abstract] [Full Text] [PDF]

				M. J. Koenigsknecht, I. Ramos, and D. M. Downs Glutamine Phosphoribosylpyrophosphate Amidotransferase-independent Phosphoribosyl Amine Synthesis from Ribose 5-Phosphate and Glutamine or Asparagine J. Biol. Chem., September 28, 2007; 282(39): 28379 - 28384. [Abstract] [Full Text] [PDF]

				M. J. Dougherty, J. M. Boyd, and D. M. Downs Inhibition of Fructose-1,6-bisphosphatase by Aminoimidazole Carboxamide Ribotide Prevents Growth of Salmonella enterica purH Mutants on Glycerol J. Biol. Chem., November 10, 2006; 281(45): 33892 - 33899. [Abstract] [Full Text] [PDF]

				M. J. Dougherty and D. M. Downs A connection between iron-sulfur cluster metabolism and the biosynthesis of 4-amino-5-hydroxymethyl-2-methylpyrimidine pyrophosphate in Salmonella enterica. Microbiology, August 1, 2006; 152(Pt 8): 2345 - 2353. [Abstract] [Full Text] [PDF]

				G. E. Schmitz and D. M. Downs An Allele of gyrA Prevents Salmonella enterica Serovar Typhimurium from Using Succinate as a Carbon Source. J. Bacteriol., April 1, 2006; 188(8): 3126 - 3129. [Abstract] [Full Text] [PDF]

				M. J. Dougherty and D. M. Downs A Mutant Allele of rpoD Results in Increased Conversion of Aminoimidazole Ribotide to Hydroxymethyl Pyrimidine in Salmonella enterica J. Bacteriol., June 15, 2004; 186(12): 4034 - 4037. [Abstract] [Full Text] [PDF]

				G. Schmitz and D. M. Downs Reduced Transaminase B (IlvE) Activity Caused by the Lack of yjgF Is Dependent on the Status of Threonine Deaminase (IlvA) in Salmonella enterica Serovar Typhimurium J. Bacteriol., February 1, 2004; 186(3): 803 - 810. [Abstract] [Full Text] [PDF]

				I. Ramos and D. M. Downs Anthranilate Synthase Can Generate Sufficient Phosphoribosyl Amine for Thiamine Synthesis in Salmonella enterica J. Bacteriol., September 1, 2003; 185(17): 5125 - 5132. [Abstract] [Full Text] [PDF]

				G. R. Bjork and K. Nilsson 1-Methylguanosine-Deficient tRNA of Salmonella enterica Serovar Typhimurium Affects Thiamine Metabolism J. Bacteriol., February 1, 2003; 185(3): 750 - 759. [Abstract] [Full Text] [PDF]

				M. Dougherty and D. M. Downs The stm4066 Gene Product of Salmonella enterica Serovar Typhimurium Has Aminoimidazole Riboside (AIRs) Kinase Activity and Allows AIRs To Satisfy the Thiamine Requirement of pur Mutant Strains J. Bacteriol., January 1, 2003; 185(1): 332 - 339. [Abstract] [Full Text] [PDF]

				S. Allen, J. L. Zilles, and D. M. Downs Metabolic Flux in Both the Purine Mononucleotide and Histidine Biosynthetic Pathways Can Influence Synthesis of the Hydroxymethyl Pyrimidine Moiety of Thiamine in Salmonella enterica J. Bacteriol., November 15, 2002; 184(22): 6130 - 6137. [Abstract] [Full Text] [PDF]

				A. Rubio and D. M. Downs Elevated Levels of Ketopantoate Hydroxymethyltransferase (PanB) Lead to a Physiologically Significant Coenzyme A Elevation in Salmonella enterica Serovar Typhimurium J. Bacteriol., May 15, 2002; 184(10): 2827 - 2832. [Abstract] [Full Text] [PDF]

				J. Gralnick and D. Downs Protection from superoxide damage associated with an increased level of the YggX protein in Salmonellla enterica PNAS, June 12, 2001; (2001) 151243198. [Abstract] [Full Text] [PDF]

				J. L. Zilles, T. J. Kappock, J. Stubbe, and D. M. Downs Altered Pathway Routing in a Class of Salmonella enterica Serovar Typhimurium Mutants Defective in Aminoimidazole Ribonucleotide Synthetase J. Bacteriol., April 1, 2001; 183(7): 2234 - 2240. [Abstract] [Full Text]

				J. L. Zilles, L. R. Croal, and D. M. Downs Action of the Thiamine Antagonist Bacimethrin on Thiamine Biosynthesis J. Bacteriol., October 1, 2000; 182(19): 5606 - 5610. [Abstract] [Full Text]

				J. Gralnick, E. Webb, B. Beck, and D. Downs Lesions in gshA (Encoding gamma -L-Glutamyl-L-Cysteine Synthetase) Prevent Aerobic Synthesis of Thiamine in Salmonella enterica Serovar Typhimurium LT2 J. Bacteriol., September 15, 2000; 182(18): 5180 - 5187. [Abstract] [Full Text]

				E. Skovran and D. M. Downs Metabolic Defects Caused by Mutations in the isc Gene Cluster in Salmonella enterica Serovar Typhimurium: Implications for Thiamine Synthesis J. Bacteriol., July 15, 2000; 182(14): 3896 - 3903. [Abstract] [Full Text]

				K. Claas, S. Weber, and D. M. Downs Lesions in the nuo Operon, Encoding NADH Dehydrogenase Complex I, Prevent PurF-Independent Thiamine Synthesis and Reduce Flux through the Oxidative Pentose Phosphate Pathway in Salmonella enterica Serovar Typhimurium J. Bacteriol., January 1, 2000; 182(1): 228 - 232. [Abstract] [Full Text]

				M. Frodyma, A. Rubio, and D. M. Downs Reduced Flux through the Purine Biosynthetic Pathway Results in an Increased Requirement for Coenzyme A in Thiamine Synthesis in Salmonella enterica Serovar Typhimurium J. Bacteriol., January 1, 2000; 182(1): 236 - 240. [Abstract] [Full Text]

				B. J. Beck and D. M. Downs A Periplasmic Location Is Essential for the Role of the ApbE Lipoprotein in Thiamine Synthesis in Salmonella typhimurium J. Bacteriol., December 1, 1999; 181(23): 7285 - 7290. [Abstract] [Full Text]

				J. L. Enos-Berlage and D. M. Downs Biosynthesis of the Pyrimidine Moiety of Thiamine Independent of the PurF Enzyme (Phosphoribosylpyrophosphate Amidotransferase) in Salmonella typhimurium: Incorporation of Stable Isotope-Labeled Glycine and Formate J. Bacteriol., February 1, 1999; 181(3): 841 - 848. [Abstract] [Full Text]

				J. L. Enos-Berlage, M. J. Langendorf, and D. M. Downs Complex Metabolic Phenotypes Caused by a Mutation in yjgF, Encoding a Member of the Highly Conserved YER057c/YjgF Family of Proteins J. Bacteriol., December 15, 1998; 180(24): 6519 - 6528. [Abstract] [Full Text]

				J. Gralnick and D. Downs Protection from superoxide damage associated with an increased level of the YggX protein in Salmonella enterica PNAS, July 3, 2001; 98(14): 8030 - 8035. [Abstract] [Full Text] [PDF]