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. 1984 Jul;4(7):1393–1401. doi: 10.1128/mcb.4.7.1393

Identification of regulatory regions within the Ty1 transposable element that regulate iso-2-cytochrome c production in the CYC7-H2 yeast mutant.

B Errede, T S Cardillo, M A Teague, F Sherman
PMCID: PMC368922  PMID: 6095068

Abstract

The CYC7-H2 mutation in the yeast Saccharomyces cerevisiae was caused by insertion of a Ty1 transposable element in front of the iso-2-cytochrome c structural gene, CYC7. The Ty1 insertion places iso-2-cytochrome c production under control of regulatory signals that are normally required for mating functions in yeast cells. We have investigated the regions of the Ty1 insertion that are responsible for the aberrant production of iso-2-cytochrome c in the CYC7-H2 mutant. Five alterations of the CYC7-H2 gene were obtained by specific restriction endonuclease cleavage of the cloned DNA and ligation of appropriate fragments. The CYC7+, CYC7-H2, and modified CYC7-H2 genes were each inserted into the yeast vector YIp5 and used to transform a cytochrome c-deficient yeast strain. Expression and regulation of each allele integrated at the CYC7 locus have been compared in vivo by determination of the amount of iso-2-cytochrome c produced. These results show that distal regions of the Ty1 element are not essential for the CYC7-H2 overproducing phenotype. In contrast, alterations in the vicinity of the proximal Ty1 junction abolish the CYC7-H2 expression and give rise to different phenotypes.

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Selected References

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

  1. Banerji J., Rusconi S., Schaffner W. Expression of a beta-globin gene is enhanced by remote SV40 DNA sequences. Cell. 1981 Dec;27(2 Pt 1):299–308. doi: 10.1016/0092-8674(81)90413-x. [DOI] [PubMed] [Google Scholar]
  2. Benoist C., Chambon P. In vivo sequence requirements of the SV40 early promotor region. Nature. 1981 Mar 26;290(5804):304–310. doi: 10.1038/290304a0. [DOI] [PubMed] [Google Scholar]
  3. Cameron J. R., Loh E. Y., Davis R. W. Evidence for transposition of dispersed repetitive DNA families in yeast. Cell. 1979 Apr;16(4):739–751. doi: 10.1016/0092-8674(79)90090-4. [DOI] [PubMed] [Google Scholar]
  4. Chaleff D. T., Fink G. R. Genetic events associated with an insertion mutation in yeast. Cell. 1980 Aug;21(1):227–237. doi: 10.1016/0092-8674(80)90130-0. [DOI] [PubMed] [Google Scholar]
  5. Ciriacy M. Cis-dominant regulatory mutations affecting the formation of glucose-repressible alcohol dehydrogenase (ADHII) in Saccharomyces cerevisiae. Mol Gen Genet. 1976 Jun 15;145(3):327–333. doi: 10.1007/BF00325831. [DOI] [PubMed] [Google Scholar]
  6. Clavilier L., Péré-Aubert G., Somlo M., Slonimski P. P. Réseau d'interactions entre des génes non liés : régulation synergique ou antagoniste de la synthèse de l'iso-1-cytochrome c, de l'iso-2-cytochrome c et du cytochrome b2. Biochimie. 1976;58(1-2):155–172. doi: 10.1016/s0300-9084(76)80366-5. [DOI] [PubMed] [Google Scholar]
  7. Clavilier L., Péré G., Slonimski P. P. Mise en évidence de plusieurs loci indépendants impliqués dans la synthèse de l'iso-2-cytochrome c chez la levure. Mol Gen Genet. 1969;104(2):195–218. doi: 10.1007/BF00272801. [DOI] [PubMed] [Google Scholar]
  8. Clewell D. B., Helinski D. R. Properties of a supercoiled deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event. Biochemistry. 1970 Oct 27;9(22):4428–4440. doi: 10.1021/bi00824a026. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Dubois E., Jacobs E., Jauniaux J. C. Expression of the ROAM mutations in Saccharomyces cerevisiae: involvement of trans-acting regulatory elements and relation with the Ty1 transcription. EMBO J. 1982;1(9):1133–1139. doi: 10.1002/j.1460-2075.1982.tb01308.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Eibel H., Philippsen P. Preferential integration of yeast transposable element Ty into a promoter region. 1984 Jan 26-Feb 1Nature. 307(5949):386–388. doi: 10.1038/307386a0. [DOI] [PubMed] [Google Scholar]
  12. Elder R. T., Loh E. Y., Davis R. W. RNA from the yeast transposable element Ty1 has both ends in the direct repeats, a structure similar to retrovirus RNA. Proc Natl Acad Sci U S A. 1983 May;80(9):2432–2436. doi: 10.1073/pnas.80.9.2432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. 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]
  16. 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]
  17. Gruss P., Dhar R., Khoury G. Simian virus 40 tandem repeated sequences as an element of the early promoter. Proc Natl Acad Sci U S A. 1981 Feb;78(2):943–947. doi: 10.1073/pnas.78.2.943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Hinnen A., Hicks J. B., Fink G. R. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. doi: 10.1073/pnas.75.4.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hsu Y. P., Kohlhaw G. B. Overproduction and control of the LEU2 gene product, beta-isopropylmalate dehydrogenase, in transformed yeast strains. J Biol Chem. 1982 Jan 10;257(1):39–41. [PubMed] [Google Scholar]
  22. Jackson J. A., Fink G. R. Gene conversion between duplicated genetic elements in yeast. Nature. 1981 Jul 23;292(5821):306–311. doi: 10.1038/292306a0. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Kingsman A. J., Gimlich R. L., Clarke L., Chinault A. C., Carbon J. Sequence variation in dispersed repetitive sequences in Saccharomyces cerevisiae. J Mol Biol. 1981 Feb 5;145(4):619–632. doi: 10.1016/0022-2836(81)90306-5. [DOI] [PubMed] [Google Scholar]
  25. Klein H. L., Petes T. D. Intrachromosomal gene conversion in yeast. Nature. 1981 Jan 15;289(5794):144–148. doi: 10.1038/289144a0. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. McKnight G. L., Cardillo T. S., Sherman F. An extensive deletion causing overproduction of yeast iso-2-cytochrome c. Cell. 1981 Aug;25(2):409–419. doi: 10.1016/0092-8674(81)90059-3. [DOI] [PubMed] [Google Scholar]
  28. Montgomery D. L., Boss J. M., McAndrew S. J., Marr L., Walthall D. A., Zitomer R. S. The molecular characterization of three transcriptional mutations in the yeast iso-2-cytochrome c gene. J Biol Chem. 1982 Jul 10;257(13):7756–7761. [PubMed] [Google Scholar]
  29. Norgard M. V., Emigholz K., Monahan J. J. Increased amplification of pBR322 plasmid deoxyribonucleic acid in Escherichia coli K-12 strains RR1 and chi1776 grown in the presence of high concentrations of nucleoside. J Bacteriol. 1979 Apr;138(1):270–272. doi: 10.1128/jb.138.1.270-272.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Parker J. H., Sherman F. Fine-structure mapping and mutational studies of gene controlling yeast cytochrome c1. Genetics. 1969 May;62(1):9–22. doi: 10.1093/genetics/62.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Roeder G. S., Fink G. R. DNA rearrangements associated with a transposable element in yeast. Cell. 1980 Aug;21(1):239–249. doi: 10.1016/0092-8674(80)90131-2. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. SHERMAN F., SLONIMSKI P. P. RESPIRATION-DEFICIENT MUTANTS OF YEAST. II. BIOCHEMISTRY. Biochim Biophys Acta. 1964 Jul 15;90:1–15. doi: 10.1016/0304-4165(64)90113-8. [DOI] [PubMed] [Google Scholar]
  36. Sherman F., Stewart J. W., Helms C., Downie J. A. Chromosome mapping of the CYC7 gene determining yeast iso-2-cytochrome c: structural and regulatory regions. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1437–1441. doi: 10.1073/pnas.75.3.1437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sherman F., Taber H., Campbell W. Genetic determination of iso-cytochromes c in yeast. J Mol Biol. 1965 Aug;13(1):21–39. doi: 10.1016/s0022-2836(65)80077-8. [DOI] [PubMed] [Google Scholar]
  38. Singh A., Sherman F. Deletions of the iso-1-cytochrome c and adjacent genes of yeast: discovery of the OSM1 gene controlling osmotic sensitivity. Genetics. 1978 Aug;89(4):653–665. doi: 10.1093/genetics/89.4.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  40. Strathern J., Hicks J., Herskowitz I. Control of cell type in yeast by the mating type locus. The alpha 1-alpha 2 hypothesis. J Mol Biol. 1981 Apr 15;147(3):357–372. doi: 10.1016/0022-2836(81)90488-5. [DOI] [PubMed] [Google Scholar]
  41. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035–1039. doi: 10.1073/pnas.76.3.1035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Szostak J. W., Wu R. Insertion of a genetic marker into the ribosomal DNA of yeast. Plasmid. 1979 Oct;2(4):536–554. doi: 10.1016/0147-619x(79)90053-2. [DOI] [PubMed] [Google Scholar]
  43. Wasylyk B., Wasylyk C., Augereau P., Chambon P. The SV40 72 bp repeat preferentially potentiates transcription starting from proximal natural or substitute promoter elements. Cell. 1983 Feb;32(2):503–514. doi: 10.1016/0092-8674(83)90470-1. [DOI] [PubMed] [Google Scholar]
  44. Williamson V. M., Cox D., Young E. T., Russell D. W., Smith M. Characterization of transposable element-associated mutations that alter yeast alcohol dehydrogenase II expression. Mol Cell Biol. 1983 Jan;3(1):20–31. doi: 10.1128/mcb.3.1.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Williamson V. M. Transposable elements in yeast. Int Rev Cytol. 1983;83:1–25. doi: 10.1016/s0074-7696(08)61684-8. [DOI] [PubMed] [Google Scholar]
  46. 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]
  47. Young T., Williamson V., Taguchi A., Smith M., Sledziewski A., Russell D., Osterman J., Denis C., Cox D., Beier D. The alcohol dehydrogenase genes of the yeast, Saccharomyces cerevisiae: isolation, structure, and regulation. Basic Life Sci. 1982;19:335–361. doi: 10.1007/978-1-4684-4142-0_26. [DOI] [PubMed] [Google Scholar]

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