• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
Mol Cell Biol. Oct 1988; 8(10): 4370–4380.
PMCID: PMC365510

Direction of chromosome rearrangements in Saccharomyces cerevisiae by use of his3 recombinational substrates.

Abstract

We used the his3 recombinational substrates (his3 fragments) to direct large interchromosomal (translocations) and intrachromosomal (deletions and tandem duplications) rearrangements in the yeast Saccharomyces cerevisiae. In strains completely deleted for the wild-type HIS3 gene, his3 fragments, one containing a deletion of 5' amino acid coding sequences and the other containing a deletion of 3' amino acid coding sequences, were first placed at preselected sites by homologous recombination. His+ revertants that arose via spontaneous mitotic recombination between the two his3 fragments were selected. This strategy was used to direct rearrangements in both RAD52+ and rad52 mutant strains. Translocations occurred in the RAD52+ genetic background and were characterized by orthogonal field alternating gel electrophoresis of yeast chromosomal DNA and by standard genetic techniques. An unexpected translocation was also identified in which HIS3 sequences were amplified. Two types of tandem duplications of the GAL(7, 10, 1) locus were also directed, and one type was not observed in rad52 mutants. Recombination mechanisms are discussed to account for these differences.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.2M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Anderson RP, Roth JR. Tandem genetic duplications in phage and bacteria. Annu Rev Microbiol. 1977;31:473–505. [PubMed]
  • Carle GF, Olson MV. Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis. Nucleic Acids Res. 1984 Jul 25;12(14):5647–5664. [PMC free article] [PubMed]
  • Carle GF, Olson MV. An electrophoretic karyotype for yeast. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3756–3760. [PMC free article] [PubMed]
  • Engels WR, Preston CR. Formation of chromosome rearrangements by P factors in Drosophila. Genetics. 1984 Aug;107(4):657–678. [PMC free article] [PubMed]
  • Fasullo MT, Davis RW. Recombinational substrates designed to study recombination between unique and repetitive sequences in vivo. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6215–6219. [PMC free article] [PubMed]
  • Fogel S, Welch JW. Tandem gene amplification mediates copper resistance in yeast. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5342–5346. [PMC free article] [PubMed]
  • Fogel S, Welch JW, Louis EJ. Meiotic gene conversion mediates gene amplification in yeast. Cold Spring Harb Symp Quant Biol. 1984;49:55–65. [PubMed]
  • Hinnen A, Hicks JB, Fink GR. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Jackson JA, Fink GR. Gene conversion between duplicated genetic elements in yeast. Nature. 1981 Jul 23;292(5821):306–311. [PubMed]
  • Johnston M, Davis RW. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Aug;4(8):1440–1448. [PMC free article] [PubMed]
  • Leblon G, Zickler D, Lebilcot S. Most Uv-Induced Reciprocal Translocations in SORDARIA MACROSPORA Occur in or near Centromere Regions. Genetics. 1986 Feb;112(2):183–204. [PMC free article] [PubMed]
  • Lehrman MA, Schneider WJ, Südhof TC, Brown MS, Goldstein JL, Russell DW. Mutation in LDL receptor: Alu-Alu recombination deletes exons encoding transmembrane and cytoplasmic domains. Science. 1985 Jan 11;227(4683):140–146. [PubMed]
  • Luria SE, Delbrück M. Mutations of Bacteria from Virus Sensitivity to Virus Resistance. Genetics. 1943 Nov;28(6):491–511. [PMC free article] [PubMed]
  • Mikus MD, Petes TD. Recombination between genes located on nonhomologous chromosomes in Saccharomyces cerevisiae. Genetics. 1982 Jul-Aug;101(3-4):369–404. [PMC free article] [PubMed]
  • Mortimer RK, Schild D. Genetic map of Saccharomyces cerevisiae. Microbiol Rev. 1980 Dec;44(4):519–571. [PMC free article] [PubMed]
  • Perkins DD. The manifestation of chromosome rearrangements in unordered asci of Neurospora. Genetics. 1974 Jul;77(3):459–489. [PMC free article] [PubMed]
  • Perkins DD, Barry EG. The cytogenetics of Neurospora. Adv Genet. 1977;19:133–285. [PubMed]
  • Potier S, Winsor B, Lacroute F. Genetic selection for reciprocal translocation at chosen chromosomal sites in Saccharomyces cerevisiae. Mol Cell Biol. 1982 Sep;2(9):1025–1032. [PMC free article] [PubMed]
  • Prakash S, Prakash L, Burke W, Montelone BA. Effects of the RAD52 Gene on Recombination in SACCHAROMYCES CEREVISIAE. Genetics. 1980 Jan;94(1):31–50. [PMC free article] [PubMed]
  • Resnick MA, 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. [PubMed]
  • Rigby PW, 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. [PubMed]
  • Roeder GS, Fink GR. DNA rearrangements associated with a transposable element in yeast. Cell. 1980 Aug;21(1):239–249. [PubMed]
  • Roman H, Fabre F. Gene conversion and associated reciprocal recombination are separable events in vegetative cells of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6912–6916. [PMC free article] [PubMed]
  • Rothstein R. Deletions of a tyrosine tRNA gene in S. cerevisiae. Cell. 1979 May;17(1):185–190. [PubMed]
  • Orr-Weaver TL, Szostak JW, Rothstein RJ. Genetic applications of yeast transformation with linear and gapped plasmids. Methods Enzymol. 1983;101:228–245. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Scherer S, Davis RW. Replacement of chromosome segments with altered DNA sequences constructed in vitro. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4951–4955. [PMC free article] [PubMed]
  • Scherer S, Davis RW. Recombination of dispersed repeated DNA sequences in yeast. Science. 1980 Sep 19;209(4463):1380–1384. [PubMed]
  • Schwartz DC, Cantor CR. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell. 1984 May;37(1):67–75. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • St John TP, Davis RW. The organization and transcription of the galactose gene cluster of Saccharomyces. J Mol Biol. 1981 Oct 25;152(2):285–315. [PubMed]
  • St John TP, Scherer S, McDonell MW, Davis RW. Deletion analysis of the Saccharomyces GAL gene cluster. Transcription from three promoters. J Mol Biol. 1981 Oct 25;152(2):317–334. [PubMed]
  • Stinchcomb DT, Mann C, Davis RW. Centromeric DNA from Saccharomyces cerevisiae. J Mol Biol. 1982 Jun 25;158(2):157–190. [PubMed]
  • Struhl K. Deletion mapping a eukaryotic promoter. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4461–4465. [PMC free article] [PubMed]
  • Struhl K. Naturally occurring poly(dA-dT) sequences are upstream promoter elements for constitutive transcription in yeast. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8419–8423. [PMC free article] [PubMed]
  • Struhl K, Davis RW. A physical, genetic and transcriptional map of the cloned his3 gene region of Saccharomyces cerevisiae. J Mol Biol. 1980 Jan 25;136(3):309–332. [PubMed]
  • Struhl K, Stinchcomb DT, Scherer S, Davis RW. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035–1039. [PMC free article] [PubMed]
  • Sugawara N, Szostak JW. Recombination between sequences in nonhomologous positions. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5675–5679. [PMC free article] [PubMed]
  • Szostak JW, Orr-Weaver TL, Rothstein RJ, Stahl FW. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. [PubMed]
  • Szostak JW, Wu R. Unequal crossing over in the ribosomal DNA of Saccharomyces cerevisiae. Nature. 1980 Apr 3;284(5755):426–430. [PubMed]
  • Willis KK, Klein HL. Intrachromosomal recombination in Saccharomyces cerevisiae: reciprocal exchange in an inverted repeat and associated gene conversion. Genetics. 1987 Dec;117(4):633–643. [PMC free article] [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...