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. 1982;1(9):1133–1139. doi: 10.1002/j.1460-2075.1982.tb01308.x

Expression of the ROAM mutations in Saccharomyces cerevisiae: involvement of trans-acting regulatory elements and relation with the Ty1 transcription.

E Dubois, E Jacobs, J C Jauniaux
PMCID: PMC553174  PMID: 6145588

Abstract

The regulatory mutations in Saccharomyces cerevisiae designated cargA + Oh, cargB + Oh, and durOh are alterations in the control regions of the respective structural genes. The alteration causing the cargA + Oh mutation has been shown to be an insertion of a Ty1 element in the 5' noncoding region of the CAR1 ( cargA ) locus. All three mutations cause overproduction of their corresponding gene products and belong to the ROAM family of mutations (Regulated Overproducing Allele responding to Mating signals) in yeast. The amount of overproduction in ROAM mutants is determined, at least in part, by signals that control mating functions in yeast. We report the identification of two genetic loci that regulate Oh mutant gene expression but that do not affect mating ability. These loci are defined by the recessive roc mutations ( ROAM mutation control) that reduce the amount of overproduction caused by the cargA + Oh, cargB + Oh, and durOh mutations. RNAs homologous to CAR1 ( cargA ), DUR1 ,2 and Ty1 DNA probes were analyzed by the Northern hybridization technique. In comparison with wild-type strains, cargA + Oh and durOh mutant strains grown on ammonia medium contain increased amounts of CAR1 and DUR1 ,2 RNA. This RNA overproduction is diminished in MATa/MAT alpha diploid strains as well as in haploid strains that also carry the ste7 mutation which prevents mating or that carry either of the roc1 or roc2 mutant alleles. The amount of RNA homologous to Ty1 DNA is also reduced in ste7 , roc1 , and roc2 mutant strains. This reduction is not observed in a strain with the ste5 mutation, which prevents mating but has no effect on overproduction of ROAM mutant gene products.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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

  1. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [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. Ciriacy M., Williamson V. M. Analysis of mutations affecting Ty-mediated gene expression in Saccharomyces cerevisiae. Mol Gen Genet. 1981;182(1):159–163. doi: 10.1007/BF00422784. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Cryer D. R., Eccleshall R., Marmur J. Isolation of yeast DNA. Methods Cell Biol. 1975;12:39–44. doi: 10.1016/s0091-679x(08)60950-4. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Deschamps J., Dubois E., Wiame J. M. L-Ornithine transaminase synthesis in Saccharomyces cerevisiae: regulation by inducer exclusion. Mol Gen Genet. 1979 Jul 24;174(3):225–232. doi: 10.1007/BF00267794. [DOI] [PubMed] [Google Scholar]
  8. Deschamps J., Wiame J. M. Mating-type effect on cis mutations leading to constitutivity of ornithine transaminase in diploid cells of Saccharomyces cerevisiae. Genetics. 1979 Jul;92(3):749–758. doi: 10.1093/genetics/92.3.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dubois E. L., Wiame J. M. Catabolic synergism: a cooperation between the availability of substrate and the need for nitrogen in the regulation of arginine catabolism in Saccharomyces cerevisiae. Mol Gen Genet. 1978 Sep 8;164(3):275–283. doi: 10.1007/BF00333157. [DOI] [PubMed] [Google Scholar]
  10. Dubois E., Grenson M., Wiame J. M. Release of the "ammonia effect" on three catabolic enzymes by NADP-specific glutamate dehydrogenaseless mutations in Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1973 Feb 20;50(4):967–972. doi: 10.1016/0006-291x(73)91500-3. [DOI] [PubMed] [Google Scholar]
  11. Dubois E., Hiernaux D., Grennon M., Wiame J. M. Specific induction of catabolism and its relation to repression of biosynthesis in arginine metabolism of Saccharomyces cerevisiae. J Mol Biol. 1978 Jul 15;122(4):383–406. doi: 10.1016/0022-2836(78)90417-5. [DOI] [PubMed] [Google Scholar]
  12. Elder R. T., St John T. P., Stinchcomb D. T., Davis R. W., Scherer S., Davis R. W. Studies on the transposable element Ty1 of yeast. I. RNA homologous to Ty1. II. Recombination and expression of Ty1 and adjacent sequences. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 2):581–591. doi: 10.1101/sqb.1981.045.01.075. [DOI] [PubMed] [Google Scholar]
  13. Errede B., Cardillo T. S., Sherman F., Dubois E., Deschamps J., Wiame J. M. Mating signals control expression of mutations resulting from insertion of a transposable repetitive element adjacent to diverse yeast genes. Cell. 1980 Nov;22(2 Pt 2):427–436. doi: 10.1016/0092-8674(80)90353-0. [DOI] [PubMed] [Google Scholar]
  14. Errede B., Cardillo T. S., Wever G., Sherman F., Stiles J. I., Friedman L. R., Sherman F. Studies on transposable elements in yeast. I. ROAM mutations causing increased expression of yeast genes: their activation by signals directed toward conjugation functions and their formation by insertion of Ty1 repetitive elements. II. deletions, duplications, and transpositions of the COR segment that encompasses the structural gene of yeast iso-1-cytochrome c. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 2):593–607. [PubMed] [Google Scholar]
  15. Farabaugh P. J., Fink G. R. Insertion of the eukaryotic transposable element Ty1 creates a 5-base pair duplication. Nature. 1980 Jul 24;286(5771):352–356. doi: 10.1038/286352a0. [DOI] [PubMed] [Google Scholar]
  16. Hartwell L. H. Mutants of Saccharomyces cerevisiae unresponsive to cell division control by polypeptide mating hormone. J Cell Biol. 1980 Jun;85(3):811–822. doi: 10.1083/jcb.85.3.811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klar A. J., Fogel S., Macleod K. MAR1-a Regulator of the HMa and HMalpha Loci in SACCHAROMYCES CEREVISIAE. Genetics. 1979 Sep;93(1):37–50. doi: 10.1093/genetics/93.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. Lemoine Y., Dubois E., Wiame J. M. The regulation of urea amidolyase of Saccharomyces cerevisiae: mating type influence on a constitutivity mutation acting in cis. Mol Gen Genet. 1978 Nov 9;166(3):251–258. [PubMed] [Google Scholar]
  20. MIDDELHOVEN W. J. THE PATHWAY OF ARGININE BREAKDOWN IN SACCHAROMYCES CEREVISIAE. Biochim Biophys Acta. 1964 Dec 9;93:650–652. doi: 10.1016/0304-4165(64)90349-6. [DOI] [PubMed] [Google Scholar]
  21. Mackay V., Manney T. R. Mutations affecting sexual conjugation and related processes in Saccharomyces cerevisiae. II. Genetic analysis of nonmating mutants. Genetics. 1974 Feb;76(2):273–288. doi: 10.1093/genetics/76.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Messenguy F., Penninckx M., Wiame J. M. Interaction between arginase and ornithine carbamoyltransferase in Saccharomyces cerevisiae. The regulatory site for ornithine. Eur J Biochem. 1971 Sep 24;22(2):277–286. doi: 10.1111/j.1432-1033.1971.tb01542.x. [DOI] [PubMed] [Google Scholar]
  24. Messenguy F. Regulation of arginine biosynthesis in Saccharomyces cerevisiae: isolation of a cis-dominant, constitutive mutant for ornithine carbamoyltransferase synthesis. J Bacteriol. 1976 Oct;128(1):49–55. doi: 10.1128/jb.128.1.49-55.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  26. Rine J., Strathern J. N., Hicks J. B., Herskowitz I. A suppressor of mating-type locus mutations in Saccharomyces cerevisiae: evidence for and identification of cryptic mating-type loci. Genetics. 1979 Dec;93(4):877–901. doi: 10.1093/genetics/93.4.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Roeder G. S., Farabaugh P. J., Chaleff D. T., Fink G. R. The origins of gene instability in yeast. Science. 1980 Sep 19;209(4463):1375–1380. doi: 10.1126/science.6251544. [DOI] [PubMed] [Google Scholar]
  28. Rothstein R. J., Sherman F. Dependence on mating type for the overproduction of iso-2-cytochrome c in the yeast mutant CYC7-H2. Genetics. 1980 Apr;94(4):891–898. doi: 10.1093/genetics/94.4.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Southern E. Gel electrophoresis of restriction fragments. Methods Enzymol. 1979;68:152–176. doi: 10.1016/0076-6879(79)68011-4. [DOI] [PubMed] [Google Scholar]
  30. Waldron C., Lacroute F. Effect of growth rate on the amounts of ribosomal and transfer ribonucleic acids in yeast. J Bacteriol. 1975 Jun;122(3):855–865. doi: 10.1128/jb.122.3.855-865.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Whitney P. A., Cooper T. G. Urea carboxylase and allophanate hydrolase. Two components of adenosine triphosphate:urea amido-lyase in Saccharomyces cerevisiae. J Biol Chem. 1972 Mar 10;247(5):1349–1353. [PubMed] [Google Scholar]
  32. Williamson V. M., Young E. T., Ciriacy M. Transposable elements associated with constitutive expression of yeast alcohol dehydrogenase II. Cell. 1981 Feb;23(2):605–614. doi: 10.1016/0092-8674(81)90156-2. [DOI] [PubMed] [Google Scholar]

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