Home LiteratureArticle Details
PMID: 3065627 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Efficient repair of HO-induced chromosomal breaks in Saccharomyces cerevisiae by recombination between flanking homologous sequences.

Molecular and cellular biology ·Vol. 8 ·No. 9 ·1988-09-00 ·Pages 3918-28

Rudin N, Haber JE

Abstract

Novel recombinational repair of a site-specific double-strand break (DSB) in a yeast chromosome was investigated. When the recognition site for the HO endonuclease enzyme is embedded in nonyeast sequences and placed between two regions of homology, expression of HO endonuclease stimulates recombination between the homologous flanking regions to yield a deletion, the apparent product of an intrachromosomal exchange between direct repeats. This deletion-repair event is very efficient, thus preventing essentially all the potential lethality due to the persistence of a DSB. Interestingly, unlike previous studies involving spontaneous recombination between chromosomal repeats, the recombination events stimulated by HO-induced DSBs are accompanied by loss of the sequences separating the homologous regions greater than 99.5% of the time. Repair is dependent on the RAD52 gene. The deletion-repair event provides an in vivo assay for the sensitivity of any particular recognition site to HO cleavage. By taking advantage of a galactose-inducible HO gene, it has been possible to follow the kinetics of this event at the DNA level and to search for intermediates in this reaction. Deletion-repair requires approximately 45 min and is inhibited when cycloheximide is added after HO endonuclease cleavage.

MeSH Terms
Chromosome Deletion Chromosomes/physiology DNA Repair Deoxyribonucleases, Type II Site-Specific/metabolism Genotype Plasmids Recombination, Genetic Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins Species Specificity
Chemicals
Saccharomyces cerevisiae Proteins HO protein, S cerevisiae SCEI protein, S cerevisiae Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Rudin N
Department of Biology, Brandeis University, Waltham, Massachusetts 02254.
Haber J E
References (45)
45 references, click to expand
  1. Deletions and single base pair changes in the yeast mating type locus that prevent homothallic mating type conversions.
    Proc Natl Acad Sci U S A. 1983 Jun;80(11):3401-5 PMID: 6304708
  2. Gene conversion between duplicated genetic elements in yeast.
    Nature. 1981 Jul 23;292(5821):306-11 PMID: 6265790
  3. Activation of Chi, a recombinator, by the action of an endonuclease at a distant site.
    Proc Natl Acad Sci U S A. 1983 Apr;80(8):2310-3 PMID: 6300909
  4. Homothallic mating type switching generates lethal chromosome breaks in rad52 strains of Saccharomyces cerevisiae.
    Mol Cell Biol. 1981 Jun;1(6):522-34 PMID: 6765605
  5. A 24-base-pair DNA sequence from the MAT locus stimulates intergenic recombination in yeast.
    Proc Natl Acad Sci U S A. 1986 Oct;83(20):7831-5 PMID: 3020559
  6. Involvement of double-strand chromosomal breaks for mating-type switching in Saccharomyces cerevisiae.
    Cold Spring Harb Symp Quant Biol. 1984;49:77-88 PMID: 6099258
  7. The repair of double-strand breaks in DNA; a model involving recombination.
    J Theor Biol. 1976 Jun;59(1):97-106 PMID: 940351
  8. Directionality of yeast mating-type interconversion.
    Cell. 1982 Mar;28(3):551-61 PMID: 7042099
  9. Effect of deletion and insertion on double-strand-break repair in Saccharomyces cerevisiae.
    Mol Cell Biol. 1987 Mar;7(3):1300-3 PMID: 3031487
  10. Rad52-independent mitotic gene conversion in Saccharomyces cerevisiae frequently results in chromosomal loss.
    Genetics. 1985 Sep;111(1):7-22 PMID: 3896928
  11. Evidence of Chromosomal Breaks near the Mating-Type Locus of SACCHAROMYCES CEREVISIAE That Accompany MATalpha xMATalpha Matings.
    Genetics. 1981 Nov;99(3-4):383-403 PMID: 17249125
  12. Yeast recombination: the association between double-strand gap repair and crossing-over.
    Proc Natl Acad Sci U S A. 1983 Jul;80(14):4417-21 PMID: 6308623
  13. The effect of cycloheximide on repair in a temperature conditional radiation-sensitive mutant of Saccharomyces cerevisiae.
    Radiat Res. 1984 Sep;99(3):582-90 PMID: 6382407
  14. Meiotic gene conversion and crossing over between dispersed homologous sequences occurs frequently in Saccharomyces cerevisiae.
    Genetics. 1987 Feb;115(2):233-46 PMID: 3549449
  15. Specific Saccharomyces cerevisiae genes are expressed in response to DNA-damaging agents.
    Mol Cell Biol. 1985 Jan;5(1):75-84 PMID: 3920512
  16. Mitotic chromosome loss in a radiation-sensitive strain of the yeast Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1981 Sep;78(9):5778-82 PMID: 7029545
  17. Mutants Showing Heterothallism from a Homothallic Strain of SACCHAROMYCES CEREVISIAE.
    Genetics. 1980 Apr;94(4):841-57 PMID: 17249021
  18. Molecular analysis of a cell lineage.
    Nature. 1983 Apr 21;302(5910):670-6 PMID: 6339953
  19. Homothallic switching of yeast mating type cassettes is initiated by a double-stranded cut in the MAT locus.
    Cell. 1982 Nov;31(1):183-92 PMID: 6297747
  20. Healing of broken linear dicentric chromosomes in yeast.
    Genetics. 1984 Feb;106(2):207-26 PMID: 6365688
  21. Regulation of RAD54- and RAD52-lacZ gene fusions in Saccharomyces cerevisiae in response to DNA damage.
    Mol Cell Biol. 1987 Mar;7(3):1078-84 PMID: 3550429
  22. One-step gene disruption in yeast.
    Methods Enzymol. 1983;101:202-11 PMID: 6310324
  23. Role of protein synthesis in the replication of yeast DNA.
    Nat New Biol. 1973 Aug 1;244(135):129-31 PMID: 4579463
  24. The product of the HO gene is a nuclease: purification and characterization of the enzyme.
    Cold Spring Harb Symp Quant Biol. 1984;49:89-96 PMID: 6099261
  25. 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):503-7 PMID: 6987653
  26. Interconversion of Yeast Mating Types I. Direct Observations of the Action of the Homothallism (HO) Gene.
    Genetics. 1976 Jun;83(2):245-58 PMID: 17248712
  27. Physical monitoring of mating type switching in Saccharomyces cerevisiae.
    Mol Cell Biol. 1988 Jun;8(6):2342-9 PMID: 2841579
  28. Effects of the RAD52 Gene on Recombination in SACCHAROMYCES CEREVISIAE.
    Genetics. 1980 Jan;94(1):31-50 PMID: 17248995
  29. Concerted deletions and inversions are caused by mitotic recombination between delta sequences in Saccharomyces cerevisiae.
    Mol Cell Biol. 1987 Mar;7(3):1198-207 PMID: 3550432
  30. Identification of a Ty insertion within the coding sequence of the S. cerevisiae URA3 gene.
    Mol Gen Genet. 1984;193(3):557-60 PMID: 6323928
  31. Double-strand breaks can initiate meiotic recombination in S. cerevisiae.
    Cell. 1986 Aug 29;46(5):733-40 PMID: 3527443
  32. Identification of a specific telomere terminal transferase activity in Tetrahymena extracts.
    Cell. 1985 Dec;43(2 Pt 1):405-13 PMID: 3907856
  33. Transfer of yeast telomeres to linear plasmids by recombination.
    Cell. 1984 Nov;39(1):191-201 PMID: 6091911
  34. Transformation of yeast with linearized plasmid DNA. Formation of inverted dimers and recombinant plasmid products.
    J Mol Biol. 1985 Aug 5;184(3):375-87 PMID: 3900413
  35. A site-specific endonuclease essential for mating-type switching in Saccharomyces cerevisiae.
    Cell. 1983 Nov;35(1):167-74 PMID: 6313222
  36. Molecular genetics of yeast mating type.
    Annu Rev Genet. 1982;16:439-500 PMID: 6760802
  37. The Role of Radiation (rad) Genes in Meiotic Recombination in Yeast.
    Genetics. 1980 Jan;94(1):51-68 PMID: 17248996
  38. A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site.
    Proc Natl Acad Sci U S A. 1982 Dec;79(23):7410-4 PMID: 6760197
  39. In vitro mutation analysis of the mating-type locus in yeast.
    Cell. 1981 Nov;27(1 Pt 2):25-35 PMID: 6276023
  40. RecBC enzyme nicking at Chi sites during DNA unwinding: location and orientation-dependence of the cutting.
    Cell. 1985 May;41(1):153-63 PMID: 3888405
  41. Yeast transformation: a model system for the study of recombination.
    Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354-8 PMID: 6273866
  42. Resolution of recombination intermediates generated during yeast mating type switching.
    Nature. 1984 Aug 30-Sep 5;310(5980):744-8 PMID: 6382020
  43. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments.
    Gene. 1979 Dec;8(1):17-24 PMID: 395030
  44. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  45. 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):119-29 PMID: 765749
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.8.9.3918-3928.1988
Published
1988-09-00
Pages
3918-28
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC365451
Subset
IM
Grants
NIGMS NIH HHS · GM07122 · United States
NIGMS NIH HHS · GM20056 · United States
External Links
PubMed source DOI: 10.1128/mcb.8.9.3918-3928.1988 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2