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Volume 153, Issue 8

Mutations in enable utilization of exogenous pyrimidine intermediates in serovar Typhimurium Free

Abstract

Mutants capable of utilizing the pyrimidine biosynthetic intermediates carbamoylaspartate and dihydroorotate for growth were derived from pyrimidine auxotrophs of serovar Typhimurium LT2. The gain-of-function phenotypes both resulted from mutations in a single gene, the third gene of a putative four-gene operon, , for which there is no homologous region in Notably, when a mutant allele was transferred to a pyrimidine-requiring strain, the transformant was then capable of using carbamoylaspartate or dihydrorotate as a pyrimidine source. The operon arrangement of the genes was supported by genetic analyses and studies employing RT-PCR, coupled to the determination of the transcriptional start site using 5′-random amplification of cDNA ends (RACE). Computer-generated predictions indicated that YhiT is an integral membrane protein with 12 putative transmembrane domains typical of bacterial transport proteins. Competition experiments showed that mutant YhiT interacts with the C-dicarboxylates succinate and malate, as well as the amino acids aspartate and asparagine. The native function of wild-type YhiT remains undetermined, but the collective results are consistent with a role as a general transporter of C-dicarboxylates and other compounds with a similar basic structure.

  • Received:
  • Accepted:
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  • Published Online:
Keyword(s): CAA, carbamoylaspartate , DHO, dihydroorotate , MNNG, N-methyl-N′-nitro-N-nitrosoguanidine , OA, orotate and RACE, random amplification of cDNA ends
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2007-08-01
2024-04-27
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Mutations in yhiT enable utilization of exogenous pyrimidine intermediates in Salmonella enterica serovar Typhimurium
Microbiology 153, 2472 (2007); https://doi.org/10.1099/mic.0.2007/007583-0
/content/journal/micro/10.1099/mic.0.2007/007583-0
/content/journal/micro/10.1099/mic.0.2007/007583-0
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