1887

Abstract

Isolation of the temperature-sensitive mutant 72c has been described previously. The mutant allele was named and causes a pleiotropic phenotype, the most striking features of which, besides temperature sensitivity, are the inability to grow on synthetic medium and supersensitivity to trimethoprim, an antibiotic that inhibits the C metabolism. This work shows that the mutation is a frameshift mutation in the gene that encodes nicotinate mononucleotide adenylyltransferase. The frameshift leads to a change of the last 10 amino acids and an addition of 17 amino acids. This lesion, renamed , leads to very little NAD and NADPH synthesis at the permissive temperature and essentially no synthesis at the non-permissive temperature. As a comparison, a new mutation in the gene, with an amino acid change in the ATP-binding site, has been isolated. Its NAD synthesis is decreased at 30 °C but the level is still sufficient to support normal growth. At 42 °C, NAD synthesis is reduced further, which leads to temperature sensitivity on minimal medium. This mutation was designated . Thus, a small decrease in NAD levels affects ability to grow on minimal medium at 42 °C, while a large decrease leads to a more pleiotropic phenotype.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.26337-0
2003-09-01
2020-08-15
Loading full text...

Full text loading...

/deliver/fulltext/micro/149/9/mic1492427.html?itemId=/content/journal/micro/10.1099/mic.0.26337-0&mimeType=html&fmt=ahah

References

  1. Bochner B. R., Ames B. N.. 1982; Complete analysis of cellular nucleotides by two-dimensional thin layer chromatography. J Biol Chem257:9759–9769
    [Google Scholar]
  2. Bullock W. O., Fernandez J. M., Short J. M.. 1987; XL1-Blue: a high efficiency plasmid transforming recA Escherichia coli strain with beta-galactosidase selection. Biotechniques5:376–379
    [Google Scholar]
  3. Chuang S.-E., Daniels D. L., Blattner F. R.. 1993; Global regulation of gene expression in Escherichia coli . J Bacteriol175:2026–2036
    [Google Scholar]
  4. Foster J. W., Moat A. G.. 1980; Nicotinamide adenine dinucleotide biosynthesis and pyridine nucleotide cycle metabolism in microbial systems. Microbiol Rev44:83–105
    [Google Scholar]
  5. Gerdes S. Y., Scholle M. D., D'Souza M.. 13 other authors 2002; From genetic footprinting to antimicrobial drug targets: examples in cofactor biosynthetic pathways. J Bacteriol184:4555–4572
    [Google Scholar]
  6. Hughes K. T., Ladika D., Roth J. R., Olivera B.. 1983a; An indispensable gene for NAD biosynthesis in Salmonella typhimurium . J Bacteriol155:213–221
    [Google Scholar]
  7. Hughes K. T., Cookson B. T., Ladika D., Olivera B. M., Roth J. R.. 1983b; 6-Aminonicotinamide-resistant mutants of Salmonella typhimurium . J Bacteriol154:1126–1136
    [Google Scholar]
  8. Isaksson L. A., Takata R.. 1978; The temperature sensitive mutant 72c. Mol Gen Genet161:9–14
    [Google Scholar]
  9. Isaksson L. A., Sköld S.-E., Skjöldebrand J., Takata R.. 1977; A procedure for isolation of spontaneous mutants with temperature sensitive synthesis of RNA and/or protein. Mol Gen Genet156:233–237
    [Google Scholar]
  10. Kurnasov O. V., Polanuyer B. M., Ananta S., Sloutsky R., Tam A., Gerdes S. Y., Osterman A. L.. 2002; Ribosylnicotinamide kinase domain of NadR protein: identification and implications in NAD biosynthesis. J Bacteriol184:6906–6917
    [Google Scholar]
  11. Lundquist R., Olivera B. M.. 1973; Pyridine nucleotide metabolism in Escherichia coli . J Biol Chem248:5137–5143
    [Google Scholar]
  12. Magni G., Amici A., Emanuelli M., Raffaelli N.. 1999; Enzymology of NAD+ synthesis. In Advances in Enzymology and Related Areas of Molecular Biology pp135–182 Edited by Purich D. L. Chichester: Wiley;
  13. Mehl R. A., Kinsland C., Begley T. P.. 2000; Identification of the Escherichia coli nicotinic acid mononucleotide adenylyltransferase gene. J Bacteriol182:4372–4374
    [Google Scholar]
  14. Miller J. H.. 1972; Formulas and recipes. In Experiments in Molecular Genetics p433 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  15. Miller J. H.. 1992; A Short Course in Bacterial Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  16. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  17. Singer M., Baker T. A., Schnitzler G.. 7 other authors 1989; A collection of strains containing genetically linked alternating antibiotic resistance elements for genetic mapping of Escherichia coli . Microbiol Rev53:1–24
    [Google Scholar]
  18. Suzuki N., Carlson J., Griffith G., Gholson R. K.. 1973; Studies on the de novo biosynthesis of NAD in Escherichia coli . V. Properties of the quinolinic acid synthetase system. Biochim Biophys Acta304:309–315
    [Google Scholar]
  19. Zhang H., Zhou T., Kurnasov O., Cheek S., Grishin N. V., Osterman A.. 2002; Crystal structures of E . coli nicotinate mononucleotide adenylyltransferase and its complex with deamido-NAD. Structure10:69–79
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.26337-0
Loading
/content/journal/micro/10.1099/mic.0.26337-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error