Skip to main content
PMC home page
Genetics logoLink to Genetics
. 1988 Jul;119(3):527–534. doi: 10.1093/genetics/119.3.527

Molecular Genetics of Serine and Threonine Catabolism in Saccharomyces Cerevisiae

JGL Petersen 1, M C Kielland-Brandt 1, T Nilsson-Tillgren 1, C Bornaes 1, S Holmberg 1
PMCID: PMC1203438  PMID: 2841185

Abstract

The catabolic L-serine (L-threonine) deaminase of Saccharomyces cerevisiae allows the yeast to grow on media with L-serine or L-threonine as sole nitrogen source. A mutant, cha1 (catabolism of hydroxyamino acids), lacking this enzyme activity has been isolated. We have cloned the CHA1 gene by complementation of a cha1 mutation. Northern analysis showed that CHA1 mRNA has a size of about 1200 ribonucleotides. CHA1 is probably the structural gene for the enzyme; it is an abundant RNA in cells grown with serine and threonine as nitrogen source, whereas it is not detected when cells are grown on ammonium or proline, i.e., the transcription of the CHA1 gene is induced by serine or threonine. Under induced growth conditions haploid ilv1 CHA1 strains do not require isoleucine, i.e., the catabolic deaminase is able to substitute for the biosynthetic threnonine deaminase encoded by the ILV1 gene. We have identified a nuclear, recessive mutation, sil1, that suppresses ilv1 mutations by increased transcription of the CHA1 gene under growth conditions leading to partial induction. The sil1 mutation could exert its effect by increasing the effective pools of the hydroxyamino acids. Alternatively SIL1 may encode a negatively acting regulatory protein for CHA1.

Full Text

The Full Text of this article is available as a PDF (2.6 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bach M. L., Lacroute F., Botstein D. Evidence for transcriptional regulation of orotidine-5'-phosphate decarboxylase in yeast by hybridization of mRNA to the yeast structural gene cloned in Escherichia coli. Proc Natl Acad Sci U S A. 1979 Jan;76(1):386–390. doi: 10.1073/pnas.76.1.386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  4. Brandriss M. C., Magasanik B. Genetics and physiology of proline utilization in Saccharomyces cerevisiae: enzyme induction by proline. J Bacteriol. 1979 Nov;140(2):498–503. doi: 10.1128/jb.140.2.498-503.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brandriss M. C. Proline utilization in Saccharomyces cerevisiae: analysis of the cloned PUT2 gene. Mol Cell Biol. 1983 Oct;3(10):1846–1856. doi: 10.1128/mcb.3.10.1846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Broach J. R., Atkins J. F., McGill C., Chow L. Identification and mapping of the transcriptional and translational products of the yeast plasmid, 2mu circle. Cell. 1979 Apr;16(4):827–839. doi: 10.1016/0092-8674(79)90098-9. [DOI] [PubMed] [Google Scholar]
  7. Cooper T. G., Lawther R. P. Induction of the allantoin degradative enzymes in Saccharomyces cerevisiae by the last intermediate of the pathway. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2340–2344. doi: 10.1073/pnas.70.8.2340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davis R. W., Thomas M., Cameron J., St John T. P., Scherer S., Padgett R. A. Rapid DNA isolations for enzymatic and hybridization analysis. Methods Enzymol. 1980;65(1):404–411. doi: 10.1016/s0076-6879(80)65051-4. [DOI] [PubMed] [Google Scholar]
  9. Feldman D. A., Datta P. Catabolite inactivation of biodegradative threonine dehydratase of Escherichia coli. Biochemistry. 1975 Apr 22;14(8):1760–1767. doi: 10.1021/bi00679a031. [DOI] [PubMed] [Google Scholar]
  10. Goss T. J., Datta P. Escherichia coli K-12 mutation that inactivates biodegradative threonine dehydratase by transposon Tn5 insertion. J Bacteriol. 1984 Jun;158(3):826–831. doi: 10.1128/jb.158.3.826-831.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goss T. J., Datta P. Molecular cloning and expression of the biodegradative threonine dehydratase gene (tdc) of Escherichia coli K12. Mol Gen Genet. 1985;201(2):308–314. doi: 10.1007/BF00425676. [DOI] [PubMed] [Google Scholar]
  12. Grenson M., Hou C., Crabeel M. Multiplicity of the amino acid permeases in Saccharomyces cerevisiae. IV. Evidence for a general amino acid permease. J Bacteriol. 1970 Sep;103(3):770–777. doi: 10.1128/jb.103.3.770-777.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grotta J., Spydell J., Pettigrew C., Ostrow P., Hunter D. The effect of nicardipine on neuronal function following ischemia. Stroke. 1986 Mar-Apr;17(2):213–219. doi: 10.1161/01.str.17.2.213. [DOI] [PubMed] [Google Scholar]
  14. Gundelach E. Suppressor studies on ilvI mutants of Saccharomyces cerevisiae. Mutat Res. 1973 Oct;20(1):25–33. doi: 10.1016/0027-5107(73)90094-8. [DOI] [PubMed] [Google Scholar]
  15. Hawthorne D C, Mortimer R K. Chromosome Mapping in Saccharomyces: Centromere-Linked Genes. Genetics. 1960 Aug;45(8):1085–1110. doi: 10.1093/genetics/45.8.1085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Isenberg S., Newman E. B. Studies on L-serine deaminase in Escherichia coli K-12. J Bacteriol. 1974 Apr;118(1):53–58. doi: 10.1128/jb.118.1.53-58.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jauniaux J. C., Dubois E., Vissers S., Crabeel M., Wiame J. M. Molecular cloning, DNA structure, and RNA analysis of the arginase gene in Saccharomyces cerevisiae. A study of cis-dominant regulatory mutations. EMBO J. 1982;1(9):1125–1131. doi: 10.1002/j.1460-2075.1982.tb01307.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. KAKAR S. N. SUPPRESSOR MUTATIONS FOR THE ISOLEUCINE LOCUS IN SACCHAROMYCES. Genetics. 1963 Aug;48:967–979. doi: 10.1093/genetics/48.8.967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kim S. S., Datta P. Chemical characterization of biodegradative threonine dehydratases from two enteric bacteria. Biochim Biophys Acta. 1982 Aug 23;706(1):27–35. doi: 10.1016/0167-4838(82)90371-5. [DOI] [PubMed] [Google Scholar]
  21. Messenguy F., Dubois E., Descamps F. Nucleotide sequence of the ARGRII regulatory gene and amino acid sequence homologies between ARGRII PPRI and GAL4 regulatory proteins. Eur J Biochem. 1986 May 15;157(1):77–81. doi: 10.1111/j.1432-1033.1986.tb09640.x. [DOI] [PubMed] [Google Scholar]
  22. Messenguy F., Dubois E. Participation of transcriptional and post-transcriptional regulatory mechanisms in the control of arginine metabolism in yeast. Mol Gen Genet. 1983;189(1):148–156. doi: 10.1007/BF00326068. [DOI] [PubMed] [Google Scholar]
  23. Middelhoven W. J. Induction and repression of arginase and ornithine transaminase in baker's yeast. Antonie Van Leeuwenhoek. 1970;36(1):1–19. doi: 10.1007/BF02069003. [DOI] [PubMed] [Google Scholar]
  24. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. doi: 10.1073/pnas.78.10.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ramos F., Wiame J. M. Occurrence of a catabolic L-serine (L-threonine) deaminase in Saccharomyces cerevisiae. Eur J Biochem. 1982 Apr;123(3):571–576. doi: 10.1111/j.1432-1033.1982.tb06570.x. [DOI] [PubMed] [Google Scholar]
  26. Saeki Y., Ito S., Shizuta Y., Hayaishi O., Kagamiyama H., Wada H. Subunit structure of biodegradative threonine deaminase. J Biol Chem. 1977 Apr 10;252(7):2206–2208. [PubMed] [Google Scholar]
  27. St John T. P., Scherer S., McDonell M. W., Davis R. W. Deletion analysis of the Saccharomyces GAL gene cluster. Transcription from three promoters. J Mol Biol. 1981 Oct 25;152(2):317–334. doi: 10.1016/0022-2836(81)90245-x. [DOI] [PubMed] [Google Scholar]
  28. Sumrada R. A., Cooper T. G. Isolation of the CAR1 gene from Saccharomyces cerevisiae and analysis of its expression. Mol Cell Biol. 1982 Dec;2(12):1514–1523. doi: 10.1128/mcb.2.12.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wang S. S., Brandriss M. C. Proline utilization in Saccharomyces cerevisiae: analysis of the cloned PUT1 gene. Mol Cell Biol. 1986 Jul;6(7):2638–2645. doi: 10.1128/mcb.6.7.2638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Whanger P. D., Phillips A. T., Rabinowitz K. W., Piperno J. R., Shada J. D., Wood W. A. The mechanism of action of 5'-adenylic acid-activated threonine dehydrase. II. Protomer-oligomer interconversions and related properties. J Biol Chem. 1968 Jan 10;243(1):167–173. [PubMed] [Google Scholar]
  31. Whiteley H. R., Tahara M. Threonine deaminase of Clostridium tetanomorphum. I. Purification and properties. J Biol Chem. 1966 Nov 10;241(21):4881–4889. [PubMed] [Google Scholar]
  32. Whitney P. A., Cooper T. G., Magasanik B. The induction of urea carboxylase and allophanate hydrolase in Saccharomyces cerevisiae. J Biol Chem. 1973 Sep 10;248(17):6203–6209. [PubMed] [Google Scholar]
  33. Whitney P. A., Magasanik B. The induction of arginase in Saccharomyces cerevisiae. J Biol Chem. 1973 Sep 10;248(17):6197–6202. [PubMed] [Google Scholar]
  34. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  35. Zimmermann F. K., Gundelach E. Intragenic complementation, hybrid enzyme formation and dominance in diploid cells of Saccharomyces cerevisiae. Mol Gen Genet. 1969;103(4):348–362. doi: 10.1007/BF00383485. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES