Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Changes in gene expression elicited by amino acid limitation in Neurospora crassa strains having normal or mutant cross-pathway amino acid control

Summary

The effects of amino acid limitation on gene expression have been investigated in Neurospora crassa strains carrying normal (cpc-1 +) or mutant (cpc-1) alleles at a locus implicated in cross-pathway amino acid control. Electrophoresis and fluorography were used to reveal the patterns of label incorporation into polypeptides in vivo, or after in vitro translation of extracted mRNAs. In a cpc-1 + strain at least 20% of detectable in vitro translation products showed relative increases in incorporation when RNA was obtained from mycelium grown under conditions of arginine limitation, by comparison with conditions of arginine sufficiency. A cpc-1 mutation, which impairs derepression of a variety of amino acid synthetic enzymes following amino acid limitation, had little detectable effect on in vivo polypeptide synthesis during amino acid sufficient growth or following pyrimidine limitation. However the mutation substantially altered the response to arginine or histidine limitation. The majority of in vitro translation products that showed increased expression in arginine limited cpc-1 + failed to increase in cpc-1 strains, but arginine limitation of cpc-1 also resulted in increases that did not occur in cpc-1 + strains. This may reflect both direct and indirect consequences of the impairment of cross-pathway control.

This is a preview of subscription content, log in to check access.

References

  1. Barthelmess IB (1982) Mutants affecting amino acid cross-pathway control in Neurospora crassa. Gen Res (Camb) 39:169–185

  2. Barthelmess IB (1984) A lethal allele at the putative regulatory locus cpc-1 of cross-pathway control in Neurospora. Mol Gen Genet 194:318–321

  3. Bode R, Casper P, Kuntze G (1983) Induction of the general control of amino acid biosynthesis in Candida spec. EH 15/D using amitrole. Biochem Physiol Pflanzen 178:457–468

  4. Bonner WM, Laskey RA (1974) A film detection method for tritium labelled proteins and nucleic acids on polyacrylamide gels. Eur J Biochem 46:83–88

  5. Carsiotis M, Jones RF (1974) Cross-pathway regulation: tryptophan mediated control of histidine and arginine biosynthetic enzymes in Neurospora crassa. J Bacteriol 119:889–892

  6. Carsiotis M, Jones RF, Wesseling M (1974) Cross-pathway regulation: histidine mediated control of histidine, tryptophan and arginine biosynthetic enzymes in Neurospora crassa. J Bacteriol 119:893–898

  7. Cramer CL, Davis RH (1984) Polyphosphate — cation interaction in the amino acid containing vacuole of Neurospora crassa. J Biol Chem 259:5152–5157

  8. Davis RH (1975) Compartmentation and regulation of fungal metabolism: genetic approaches. Annu Rev Genet 9:39–67

  9. Davis RH (1979) Genetics of arginine biosynthesis in Neurospora crassa. Genetics 93:557–575

  10. Davis RH, Ristow JL, Ginsburgh CL (1981) Independent localization and regulation of carbamyl phosphate synthetase A polypeptides of Neurospora crassa. Mol Gen Genet 181:215–221

  11. Delforge J, Messenguy F, Wiame J-M (1975) The regulation of arginine biosynthesis in Saccharomyces cerevisiae. The specificity of arg R mutations and the general control of amino acid biosynthesis. Eur J Biochem 57:231–239

  12. Flint HJ, Kemp BF (1981) General control of arginine biosynthetic enzymes in Neurospora crassa. J Gen Microbiol 124:129–140

  13. Flint HJ, Porteous DJ, Kacser H (1980) Control of the flux in the arginine pathway of Neurospora crassa: the flux from citrulline to arginine. Biochem J 190:1–15

  14. Gallant JA (1979) Stringent control in E. coli. Annu Rev Genet 13:393–415

  15. Hinnebusch AG, Fink GR (1983) Positive regulation in the general amino acid control of Saccharomyces cerevisiae. Proc Natl Acad Sci USA 80:5374–5378

  16. Jones EW, Fink GR (1982) Regulation of amino acid and nucleotide biosynthesis in yeast. In: Strathern JN, Jones EW and Broach JR (eds) Molecular biology of the yeast Saccharomyces cerevisiae — metabolism and gene expression. Cold Spring Harbor Laboratory, New York, pp 181–287

  17. Kinnaird JH, Keighren MA, Kinsey JA, Eaton, M, Fincham JRS (1982) Cloning of the am (glutamate dehydrogenase) gene of Neurospora crassa through the use of a synthetic DNA probe. Gene 20:387–396

  18. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

  19. Niederberger P, Miozzari G, Hütter R (1981) Biological role of the general control of amino acid biosynthesis in Saccharomyces cerevisiae. Mol Cell Biol 1:584–593

  20. Niederberger P, Aebi M, Hütter R (1983) Influence of the general control of amino acid biosynthesis on cell growth and viability in Saccharomyces cerevisiae. J Gen Microbiol 129:2571–2583

  21. O'Farrell PH (1975) High resolution two dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021

  22. O'Farrell PH (1978) The suppression of defective translation by ppGpp and its role in the stringent response. Cell 14:545–557

  23. Penn MD, Galgoci B, Greer H (1983) Identification of AAS genes and their regulatory role in general control of amino acid biosynthesis in yeast. Proc Natl Acad Sci USA 80:2704–2708

  24. Perkins DD, Radford A, Newmeyer D, Bjorkman M (1982) Chromosomal loci of Neurospora crassa. Microbiol Rev 46:426–570

  25. Piotrowska M (1980) Cross-pathway regulation of ornithine carbamoyl transferase synthesis in Aspergillus nidulans. J Gen Microbiol 116:335–339

  26. Schürch A, Miozzari G, Hütter R (1974) Regulation of tryptophan biosynthesis in Saccharomyces cerevisiae: mode of action of 5 methyl tryptophan and 5 methyl tryptophan sensitive mutatns. J Bacteriol 117:1131–1140

  27. Shulman RW, Sripati CE, Warner JR (1977) Non-coordinated transcription in the absence of protein synthesis in yeast. J Biol Chem 252:1344–1349

  28. Vogel HJ (1956) A convenient growth medium for Neurospora (medium N). Microbiol Genet Bull 13:42–43

  29. Warner JR, Gorenstein C (1978) Yeast has a true stringent response. Nature 275:338–339

Download references

Author information

Additional information

Communicated by C. Auerbach

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Flint, H.J. Changes in gene expression elicited by amino acid limitation in Neurospora crassa strains having normal or mutant cross-pathway amino acid control. Molec. Gen. Genet. 200, 283–290 (1985). https://doi.org/10.1007/BF00425437

Download citation

Keywords

  • Electrophoresis
  • Polypeptide
  • Arginine
  • Histidine
  • Pyrimidine