Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 1988 Sep;120(1):57–62. doi: 10.1093/genetics/120.1.57

A Highly Revertible Cyc1 Mutant of Yeast Contains a Small Tandem Duplication

G Das 1, S Consaul 1, F Sherman 1
PMCID: PMC1203505  PMID: 2851481

Abstract

A mutant, cyc1-96, that reverts spontaneously at an extremely high rate, was uncovered after examining approximately 500 cyc1 mutants which lack or have defective iso-1-cytochrome c in the yeast Saccharomyces cerevisiae. Cloning and DNA sequencing of appropriate fragments revealed that the cyc1-96 mutation contained a 19 bp duplication whereas the spontaneously arising revertants contained the normal wild-type sequence. Because the 19 bp segment in the wild-type sequence is flanked by a 5 bp repeat and because the cyc1-96 mutation arose spontaneously, the 19 bp duplication may have arisen by slippage and misalignment during DNA synthesis. The high reversion rate was not diminished in strains containing the rad52 mutation, which generally reduces mitotic recombination, including recombination associated with the elimination of a segment of a long direct repeat. Thus the loss of segments from short and long duplications occur by different mechanisms. We suggest that the high reversion rates of cyc1-96 and other short duplications are due to misalignment errors during replication.

Full Text

The Full Text of this article is available as a PDF (1.8 MB).

Selected References

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

  1. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Borts R. H., Lichten M., Haber J. E. Analysis of meiosis-defective mutations in yeast by physical monitoring of recombination. Genetics. 1986 Jul;113(3):551–567. doi: 10.1093/genetics/113.3.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Calos M. P., Galas D., Miller J. H. Genetic studies of the lac repressor. VIII. DNA sequence change resulting from an intragenic duplication. J Mol Biol. 1978 Dec 25;126(4):865–869. doi: 10.1016/0022-2836(78)90025-6. [DOI] [PubMed] [Google Scholar]
  4. Efstratiadis A., Posakony J. W., Maniatis T., Lawn R. M., O'Connell C., Spritz R. A., DeRiel J. K., Forget B. G., Weissman S. M., Slightom J. L. The structure and evolution of the human beta-globin gene family. Cell. 1980 Oct;21(3):653–668. doi: 10.1016/0092-8674(80)90429-8. [DOI] [PubMed] [Google Scholar]
  5. Ernst J. F., Stewart J. W., Sherman F. Formation of composite iso-cytochromes c by recombination between non-allelic genes of yeast. J Mol Biol. 1982 Nov 5;161(3):373–394. doi: 10.1016/0022-2836(82)90245-5. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Farabaugh P. J., Schmeissner U., Hofer M., Miller J. H. Genetic studies of the lac repressor. VII. On the molecular nature of spontaneous hotspots in the lacI gene of Escherichia coli. J Mol Biol. 1978 Dec 25;126(4):847–857. doi: 10.1016/0022-2836(78)90023-2. [DOI] [PubMed] [Google Scholar]
  8. Haber J. E., Hearn M. Rad52-independent mitotic gene conversion in Saccharomyces cerevisiae frequently results in chromosomal loss. Genetics. 1985 Sep;111(1):7–22. doi: 10.1093/genetics/111.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hampsey D. M., Das G., Sherman F. Amino acid replacements in yeast iso-1-cytochrome c. Comparison with the phylogenetic series and the tertiary structure of related cytochromes c. J Biol Chem. 1986 Mar 5;261(7):3259–3271. [PubMed] [Google Scholar]
  10. Hampsey D. M., Das G., Sherman F. Yeast iso-1-cytochrome c: genetic analysis of structural requirements. FEBS Lett. 1988 Apr 25;231(2):275–283. doi: 10.1016/0014-5793(88)80834-2. [DOI] [PubMed] [Google Scholar]
  11. Hampsey D. M., Ernst J. F., Stewart J. W., Sherman F. Multiple base-pair mutations in yeast. J Mol Biol. 1988 Jun 5;201(3):471–486. doi: 10.1016/0022-2836(88)90629-8. [DOI] [PubMed] [Google Scholar]
  12. Hampsey D. M., Koski R. A., Sherman F. Highly mutable sites for ICR-170-induced frameshift mutations are associated with potential DNA hairpin structures: studies with SUP4 and other Saccharomyces cerevisiae genes. Mol Cell Biol. 1986 Dec;6(12):4425–4432. doi: 10.1128/mcb.6.12.4425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Liebman S. W., Singh A., Sherman F. A mutator affecting the region of the iso-1-cytochrome c gene in yeast. Genetics. 1979 Jul;92(3):783–802. doi: 10.1093/genetics/92.3.783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Liebman S., Shalit P., Picologlou S. Ty elements are involved in the formation of deletions in DEL1 strains of Saccharomyces cerevisiae. Cell. 1981 Nov;26(3 Pt 1):401–409. doi: 10.1016/0092-8674(81)90209-9. [DOI] [PubMed] [Google Scholar]
  15. Loprieno N., Abbondandolo A., Bonatti S., Guglielminetti R. Analysis of the genetic instability induced by nitrous acid in Schizosaccharomyces pombe. Genet Res. 1968 Aug;12(1):45–54. doi: 10.1017/s0016672300011605. [DOI] [PubMed] [Google Scholar]
  16. Malone R. E., Esposito R. E. Recombinationless meiosis in Saccharomyces cerevisiae. Mol Cell Biol. 1981 Oct;1(10):891–901. doi: 10.1128/mcb.1.10.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Malone R. E., Esposito R. E. The RAD52 gene is required for homothallic interconversion of mating types and spontaneous mitotic recombination in yeast. Proc Natl Acad Sci U S A. 1980 Jan;77(1):503–507. doi: 10.1073/pnas.77.1.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moore C. W., Hampsey D. M., Ernst J. F., Sherman F. Differential mismatch repair can explain the disproportionalities between physical distances and recombination frequencies of cyc1 mutations in yeast. Genetics. 1988 May;119(1):21–34. doi: 10.1093/genetics/119.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mortimer R. K., Johnston J. R. Genealogy of principal strains of the yeast genetic stock center. Genetics. 1986 May;113(1):35–43. doi: 10.1093/genetics/113.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nasim A., Grant C. Genetic analysis of replicating instabilities in yeast. Mutat Res. 1973 Feb;17(2):185–190. doi: 10.1016/0027-5107(73)90165-6. [DOI] [PubMed] [Google Scholar]
  21. Nasim A., Stephen E. R., Erratt J. A. Replicating instabilities in yeast: occurrence in different mutational systems. Mol Gen Genet. 1985;199(1):152–153. doi: 10.1007/BF00327525. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Rambosek J. A., Kinsey J. A. An unstable mutant gene of the am locus of Neurospora results from a small duplication. Gene. 1984 Jan;27(1):101–107. doi: 10.1016/0378-1119(84)90242-7. [DOI] [PubMed] [Google Scholar]
  24. Ramdas L., Sherman F., Nall B. T. Guanidine hydrochloride induced equilibrium unfolding of mutant forms of iso-1-cytochrome c with replacement of proline-71. Biochemistry. 1986 Nov 4;25(22):6952–6958. doi: 10.1021/bi00370a032. [DOI] [PubMed] [Google Scholar]
  25. Resnick M. A., Martin P. The repair of double-strand breaks in the nuclear DNA of Saccharomyces cerevisiae and its genetic control. Mol Gen Genet. 1976 Jan 16;143(2):119–129. doi: 10.1007/BF00266917. [DOI] [PubMed] [Google Scholar]
  26. Rothstein R., Helms C., Rosenberg N. Concerted deletions and inversions are caused by mitotic recombination between delta sequences in Saccharomyces cerevisiae. Mol Cell Biol. 1987 Mar;7(3):1198–1207. doi: 10.1128/mcb.7.3.1198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schweingruber M. E., Stewart J. W., Sherman F. Primary site and second site revertants of missense mutants of the evolutionarily invariant tryptophan 64 in iso-1-cytochrome c from yeast. J Biol Chem. 1979 May 25;254(10):4132–4143. [PubMed] [Google Scholar]
  28. Sherman F., Jackson M., Liebman S. W., Schweingruber A. M., Stewart J. W. A deletion map of cyc1 mutants and its correspondence to mutationally altered iso-1-cytochromes c of yeast. Genetics. 1975 Sep;81(1):51–73. doi: 10.1093/genetics/81.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sherman F., Stewart J. W., Jackson M., Gilmore R. A., Parker J. H. Mutants of yeast defective in iso-1-cytochrome c. Genetics. 1974 Jun;77(2):255–284. doi: 10.1093/genetics/77.2.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Smith A. J. DNA sequence analysis by primed synthesis. Methods Enzymol. 1980;65(1):560–580. doi: 10.1016/s0076-6879(80)65060-5. [DOI] [PubMed] [Google Scholar]
  31. Stiles J. I., Friedman L. R., Helms C., Consaul S., Sherman F. Transposition of the gene cluster CYC1-OSM1-RAD7 in yeast. J Mol Biol. 1981 Jun 5;148(4):331–346. doi: 10.1016/0022-2836(81)90179-0. [DOI] [PubMed] [Google Scholar]
  32. Streisinger G., Okada Y., Emrich J., Newton J., Tsugita A., Terzaghi E., Inouye M. Frameshift mutations and the genetic code. This paper is dedicated to Professor Theodosius Dobzhansky on the occasion of his 66th birthday. Cold Spring Harb Symp Quant Biol. 1966;31:77–84. doi: 10.1101/sqb.1966.031.01.014. [DOI] [PubMed] [Google Scholar]
  33. Weiffenbach B., Haber J. E. Homothallic mating type switching generates lethal chromosome breaks in rad52 strains of Saccharomyces cerevisiae. Mol Cell Biol. 1981 Jun;1(6):522–534. doi: 10.1128/mcb.1.6.522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Winston F., Chaleff D. T., Valent B., Fink G. R. Mutations affecting Ty-mediated expression of the HIS4 gene of Saccharomyces cerevisiae. Genetics. 1984 Jun;107(2):179–197. doi: 10.1093/genetics/107.2.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zaret K. S., Sherman F. DNA sequence required for efficient transcription termination in yeast. Cell. 1982 Mar;28(3):563–573. doi: 10.1016/0092-8674(82)90211-2. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES