@article{mbs:/content/journal/micro/10.1099/13500872-142-2-359, author = "Post, David A. and Switzer, Robert L. and Hove-Jensen, Bjarne", title = "The defective phosphoribosyl diphosphate synthase in a temperature-sensitive prs-2 mutant of Escherichia coli is compensated by increased enzyme synthesis", journal= "Microbiology", year = "1996", volume = "142", number = "2", pages = "359-365", doi = "https://doi.org/10.1099/13500872-142-2-359", url = "https://www.microbiologyresearch.org/content/journal/micro/10.1099/13500872-142-2-359", publisher = "Microbiology Society", issn = "1465-2080", type = "Journal Article", keywords = "RNA analysis", keywords = "nucleotide synthesis", keywords = "PRPP synthase", keywords = "phosphoribosylpyrophosphate", keywords = "prs gene", abstract = "Summary: An Escherichia coli strain which is temperature-sensitive for growth due to a mutation(prs-2)causing a defective phosphoribosyl diphosphate(PRPP)synthase has been characterized. The temperature-sensitive mutation was mapped to a 276 bp HindIII-BssHII DNA fragment located within the open reading frame specifying the PRPP synthase polypeptide. Cloning and sequencing of the mutant allele revealed two mutations. One, a G→A transition, located in the ninth codon, was responsible for the temperature-conditional phenotype and resulted in a serine residue at this position. The wild-type codon at this position specified a glycine residue that is conserved among PRPP synthases across a broad phylogenetic range. Cells harbouring the glycine-to-serine alteration specified by a plasmid contained approximately 50% of the PRPP synthase activity of cells harbouring a plasmid-borne wildtype allele, both grown at 25°C. The mutant enzyme had nearly normal heat stability, as long as it was synthesized at 25°C. In contrast, there was hardly any PRPP synthase activity or anti-PRPP synthase antibody cross-reactive material present in cells harbouring the glycine to serine alteration following temperature shift to 42°C. The other mutation was aC→T transition located 39 bp upstream of the G→A mutation, i.e. outside the coding sequence and close to the Shine-Dalgarno sequence. Cells harbouring only the C→T mutation in a plasmid contained approximately three times as much PRPP synthase activity as a strain harbouring a plasmid-borne wild-type prs allele. In cells harbouring both mutations, the C→T mutation appeared to compensate for the G→A mutation by increasing the amount of a partially defective enzyme at the permissive temperature.", }