Home LiteratureArticle Details
PMID: 10409756 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Stability of the human fragile X (CGG)(n) triplet repeat array in Saccharomyces cerevisiae deficient in aspects of DNA metabolism.

Molecular and cellular biology ·Vol. 19 ·No. 8 ·1999-08-00 ·Pages 5675-84

White PJ, Borts RH, Hirst MC

Abstract

Expanded trinucleotide repeats underlie a growing number of human diseases. The human FMR1 (CGG)(n) array can exhibit genetic instability characterized by progressive expansion over several generations leading to gene silencing and the development of the fragile X syndrome. While expansion is dependent upon the length of uninterrupted (CGG)(n), instability occurs in a limited germ line and early developmental window, suggesting that lineage-specific expression of other factors determines the cellular environment permissive for expansion. To identify these factors, we have established normal- and premutation-length human FMR1 (CGG)(n) arrays in the yeast Saccharomyces cerevisiae and assessed the frequency of length changes greater than 5 triplets in cells deficient in various DNA repair and replication functions. In contrast to previous studies with Escherichia coli, we observed a low frequency of orientation-dependent large expansions in arrays carrying long uninterrupted (CGG)(n) arrays in a wild-type background. This frequency was unaffected by deletion of several DNA mismatch repair genes or deletion of the EXO1 and DIN7 genes and was not enhanced through meiosis in a wild-type background. Array contraction occurred in an orientation-dependent manner in most mutant backgrounds, but loss of the Sgs1p resulted in a generalized increase in array stability in both orientations. In contrast, FMR1 arrays had a 10-fold-elevated frequency of expansion in a rad27 background, providing evidence for a role in lagging-strand Okazaki fragment processing in (CGG)(n) triplet repeat expansion.

MeSH Terms
14-3-3 Proteins Checkpoint Kinase 1 DNA Helicases/deficiency,genetics,physiology DNA Repair/genetics DNA, Fungal/metabolism DNA, Recombinant/chemistry,metabolism Escherichia coli/genetics,metabolism Exodeoxyribonucleases Fragile X Mental Retardation Protein Fragile X Syndrome/genetics Fungal Proteins/genetics,physiology Humans Models, Genetic Nerve Tissue Proteins/genetics Nucleic Acid Conformation Protein Kinases/deficiency,genetics,physiology Proteins/genetics,physiology RNA-Binding Proteins RecQ Helicases Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Trinucleotide Repeat Expansion Trinucleotide Repeats Tyrosine 3-Monooxygenase
Chemicals
14-3-3 Proteins DNA, Fungal DNA, Recombinant FMR1 protein, human Fungal Proteins Nerve Tissue Proteins Proteins RNA-Binding Proteins Saccharomyces cerevisiae Proteins Fragile X Mental Retardation Protein Tyrosine 3-Monooxygenase Protein Kinases CHEK1 protein, human Checkpoint Kinase 1 DIN7 protein, S cerevisiae Exodeoxyribonucleases SGS1 protein, S cerevisiae DNA Helicases RecQ Helicases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
White P J
Fragile X Group, Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Headington, Oxford OX3 9DS, United Kingdom.
Borts R H
Hirst M C
References (72)
72 references, click to expand
  1. Length of uninterrupted CGG repeats determines instability in the FMR1 gene.
    Nat Genet. 1994 Sep;8(1):88-94 PMID: 7987398
  2. Characterization of a mutant strain of Saccharomyces cerevisiae with a deletion of the RAD27 gene, a structural homolog of the RAD2 nucleotide excision repair gene.
    J Bacteriol. 1995 Jan;177(2):364-71 PMID: 7814325
  3. Sequence analysis of the fragile X trinucleotide repeat: implications for the origin of the fragile X mutation.
    Hum Mol Genet. 1994 Sep;3(9):1543-51 PMID: 7833909
  4. Precursor arrays for triplet repeat expansion at the fragile X locus.
    Hum Mol Genet. 1994 Sep;3(9):1553-60 PMID: 7833910
  5. Conditional lethality of null mutations in RTH1 that encodes the yeast counterpart of a mammalian 5'- to 3'-exonuclease required for lagging strand DNA synthesis in reconstituted systems.
    J Biol Chem. 1995 Mar 3;270(9):4193-6 PMID: 7876174
  6. Stability of an expanded trinucleotide repeat in the androgen receptor gene in transgenic mice.
    Nat Genet. 1995 Feb;9(2):191-6 PMID: 7719348
  7. The fragile X premutation in carriers and its effect on mutation size in offspring.
    Am J Hum Genet. 1995 May;56(5):1147-55 PMID: 7726171
  8. New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae.
    Yeast. 1994 Dec;10(13):1793-808 PMID: 7747518
  9. The FEN-1 family of structure-specific nucleases in eukaryotic DNA replication, recombination and repair.
    Bioessays. 1997 Mar;19(3):233-40 PMID: 9080773
  10. Trinucleotide repeats associated with human disease.
    Nucleic Acids Res. 1997 Jun 15;25(12):2245-54 PMID: 9171073
  11. Repeat expansion--all in a flap?
    Nat Genet. 1997 Jun;16(2):116-8 PMID: 9171819
  12. Identification and characterization of Saccharomyces cerevisiae EXO1, a gene encoding an exonuclease that interacts with MSH2.
    Proc Natl Acad Sci U S A. 1997 Jul 8;94(14):7487-92 PMID: 9207118
  13. Second pathway for completion of human DNA base excision-repair: reconstitution with purified proteins and requirement for DNase IV (FEN1).
    EMBO J. 1997 Jun 2;16(11):3341-8 PMID: 9214649
  14. Transition from premutation to full mutation in fragile X syndrome is likely to be prezygotic.
    Hum Mol Genet. 1997 Jul;6(7):971-9 PMID: 9215664
  15. Human MSH2 binds to trinucleotide repeat DNA structures associated with neurodegenerative diseases.
    Hum Mol Genet. 1997 Jul;6(7):1117-23 PMID: 9215683
  16. Trinucleotide repeats (CGG)22TGG(CGG)43TGG(CGG)21 from the fragile X gene remain stable in transgenic mice.
    Hum Genet. 1997 Sep;100(3-4):407-14 PMID: 9272164
  17. Trinucleotide repeats affect DNA replication in vivo.
    Nat Genet. 1997 Nov;17(3):298-304 PMID: 9354793
  18. Expansions of CAG repeat tracts are frequent in a yeast mutant defective in Okazaki fragment maturation.
    Hum Mol Genet. 1998 Jan;7(1):69-74 PMID: 9384605
  19. Dynamic mutation: possible mechanisms and significance in human disease.
    Trends Biochem Sci. 1997 Nov;22(11):432-6 PMID: 9397685
  20. FMR1 premutation allele (CGG)81 is stable in mice.
    Eur J Hum Genet. 1997 Sep-Oct;5(5):293-8 PMID: 9412786
  21. Expansion and length-dependent fragility of CTG repeats in yeast.
    Science. 1998 Feb 6;279(5352):853-6 PMID: 9452383
  22. Unusual mutations in high functioning fragile X males: apparent instability of expanded unmethylated CGG repeats.
    J Med Genet. 1998 Feb;35(2):103-11 PMID: 9507388
  23. The yeast HSM3 gene acts in one of the mismatch repair pathways.
    Genetics. 1998 Mar;148(3):963-73 PMID: 9539417
  24. Destabilization of yeast micro- and minisatellite DNA sequences by mutations affecting a nuclease involved in Okazaki fragment processing (rad27) and DNA polymerase delta (pol3-t).
    Mol Cell Biol. 1998 May;18(5):2779-88 PMID: 9566897
  25. Cloned human FMR1 trinucleotide repeats exhibit a length- and orientation-dependent instability suggestive of in vivo lagging strand secondary structure.
    Nucleic Acids Res. 1998 May 15;26(10):2353-8 PMID: 9580685
  26. Hex1: a new human Rad2 nuclease family member with homology to yeast exonuclease 1.
    Nucleic Acids Res. 1998 Aug 15;26(16):3762-8 PMID: 9685493
  27. Mismatch repair-induced meiotic recombination requires the pms1 gene product.
    Genetics. 1990 Mar;124(3):573-84 PMID: 2179055
  28. Instability of a 550-base pair DNA segment and abnormal methylation in fragile X syndrome.
    Science. 1991 May 24;252(5009):1097-102 PMID: 2031184
  29. Fragile X genotype characterized by an unstable region of DNA.
    Science. 1991 May 24;252(5009):1179-81 PMID: 2031189
  30. Identification of a gene (FMR-1) containing a CGG repeat coincident with a breakpoint cluster region exhibiting length variation in fragile X syndrome.
    Cell. 1991 May 31;65(5):905-14 PMID: 1710175
  31. Absence of expression of the FMR-1 gene in fragile X syndrome.
    Cell. 1991 Aug 23;66(4):817-22 PMID: 1878973
  32. Variation of the CGG repeat at the fragile X site results in genetic instability: resolution of the Sherman paradox.
    Cell. 1991 Dec 20;67(6):1047-58 PMID: 1760838
  33. Genotype mosaicism in fragile X fetal tissues.
    Hum Genet. 1992 Apr;89(1):114-6 PMID: 1349561
  34. Instability of simple sequence DNA in Saccharomyces cerevisiae.
    Mol Cell Biol. 1992 Jun;12(6):2749-57 PMID: 1588966
  35. Analysis of full fragile X mutations in fetal tissues and monozygotic twins indicate that abnormal methylation and somatic heterogeneity are established early in development.
    Am J Med Genet. 1992 Apr 15-May 1;43(1-2):208-16 PMID: 1605193
  36. Inheritance of the fragile X syndrome: size of the fragile X premutation is a major determinant of the transition to full mutation.
    J Med Genet. 1992 Nov;29(11):794-801 PMID: 1453430
  37. Bloom's syndrome. XVIII. Hypermutability at a tandem-repeat locus.
    Hum Genet. 1992 Dec;90(4):360-7 PMID: 1483691
  38. Methylation analysis of CGG sites in the CpG island of the human FMR1 gene.
    Hum Mol Genet. 1992 Nov;1(8):571-8 PMID: 1301165
  39. DNA methylation represses FMR-1 transcription in fragile X syndrome.
    Hum Mol Genet. 1992 Sep;1(6):397-400 PMID: 1301913
  40. Mitotic stability of fragile X mutations in differentiated cells indicates early post-conceptional trinucleotide repeat expansion.
    Nat Genet. 1993 Jun;4(2):140-2 PMID: 8348151
  41. The full mutation in the FMR-1 gene of male fragile X patients is absent in their sperm.
    Nat Genet. 1993 Jun;4(2):143-6 PMID: 8348152
  42. Destabilization of tracts of simple repetitive DNA in yeast by mutations affecting DNA mismatch repair.
    Nature. 1993 Sep 16;365(6443):274-6 PMID: 8371783
  43. Simple repeat DNA is not replicated simply.
    Nat Genet. 1994 Feb;6(2):114-6 PMID: 8162063
  44. CGG repeats associated with DNA instability and chromosome fragility form structures that block DNA synthesis in vitro.
    Nucleic Acids Res. 1995 Oct 25;23(20):4202-9 PMID: 7479085
  45. Stability of microsatellites and minisatellites in Bloom syndrome, a human syndrome of genetic instability.
    Mutat Res. 1996 Apr 2;362(3):227-36 PMID: 8637501
  46. Cloning, characterization and properties of plasmids containing CGG triplet repeats from the FMR-1 gene.
    J Mol Biol. 1996 May 17;258(4):614-26 PMID: 8636996
  47. Biochemistry and genetics of eukaryotic mismatch repair.
    Genes Dev. 1996 Jun 15;10(12):1433-42 PMID: 8666228
  48. Nucleosome assembly on CTG triplet repeats.
    J Biol Chem. 1996 Jun 21;271(25):15222-9 PMID: 8663027
  49. Trinucleotide repeats that expand in human disease form hairpin structures in vitro.
    Cell. 1995 May 19;81(4):533-40 PMID: 7758107
  50. Sequence of human FEN-1, a structure-specific endonuclease, and chromosomal localization of the gene (FEN1) in mouse and human.
    Genomics. 1995 Jan 1;25(1):220-5 PMID: 7774922
  51. Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure.
    Yeast. 1995 Apr 15;11(4):355-60 PMID: 7785336
  52. Expanded CTG triplet blocks from the myotonic dystrophy gene create the strongest known natural nucleosome positioning elements.
    Genomics. 1995 Jan 20;25(2):570-3 PMID: 7789994
  53. Requirement of the yeast RTH1 5' to 3' exonuclease for the stability of simple repetitive DNA.
    Science. 1995 Jul 14;269(5221):238-40 PMID: 7618086
  54. Fragile X gene instability: anchoring AGGs and linked microsatellites.
    Am J Hum Genet. 1995 Aug;57(2):351-61 PMID: 7668261
  55. Mechanisms of DNA expansion.
    Chromosoma. 1995 Oct;104(1):2-13 PMID: 7587591
  56. Pausing of DNA synthesis in vitro at specific loci in CTG and CGG triplet repeats from human hereditary disease genes.
    J Biol Chem. 1995 Nov 10;270(45):27014-21 PMID: 7592950
  57. SGS1, a homologue of the Bloom's and Werner's syndrome genes, is required for maintenance of genome stability in Saccharomyces cerevisiae.
    Genetics. 1996 Nov;144(3):935-45 PMID: 8913739
  58. Orientation dependence of trinucleotide CAG repeat instability in Saccharomyces cerevisiae.
    Mol Cell Biol. 1996 Dec;16(12):6617-22 PMID: 8943315
  59. Mutator specificity and disease: looking over the FENce.
    Cell. 1997 Jan 24;88(2):155-8 PMID: 9008154
  60. A novel mutation avoidance mechanism dependent on S. cerevisiae RAD27 is distinct from DNA mismatch repair.
    Cell. 1997 Jan 24;88(2):253-63 PMID: 9008166
  61. Characterization of the full fragile X syndrome mutation in fetal gametes.
    Nat Genet. 1997 Feb;15(2):165-9 PMID: 9020841
  62. Moderate intergenerational and somatic instability of a 55-CTG repeat in transgenic mice.
    Nat Genet. 1997 Feb;15(2):190-2 PMID: 9020847
  63. Hypermutable myotonic dystrophy CTG repeats in transgenic mice.
    Nat Genet. 1997 Feb;15(2):193-6 PMID: 9020848
  64. Instability of highly expanded CAG repeats in mice transgenic for the Huntington's disease mutation.
    Nat Genet. 1997 Feb;15(2):197-200 PMID: 9020849
  65. A yeast replicative helicase, Dna2 helicase, interacts with yeast FEN-1 nuclease in carrying out its essential function.
    Mol Cell Biol. 1997 Apr;17(4):2136-42 PMID: 9121462
  66. Tosca: a Drosophila gene encoding a nuclease specifically expressed in the female germline.
    Dev Biol. 1996 Aug 25;178(1):90-100 PMID: 8812111
  67. Destabilization of CAG trinucleotide repeat tracts by mismatch repair mutations in yeast.
    Hum Mol Genet. 1997 Mar;6(3):349-55 PMID: 9147637
  68. Characterization of a novel DNA damage-inducible gene of Saccharomyces cerevisiae, DIN7, which is a structural homolog of the RAD2 and RAD27 DNA repair genes.
    Mol Gen Genet. 1997 Feb 27;253(6):655-65 PMID: 9079876
  69. In vitro expansion of GGC:GCC repeats: identification of the preferred strand of expansion.
    Nucleic Acids Res. 1996 Jul 15;24(14):2835-40 PMID: 8759019
  70. Nucleosome assembly on methylated CGG triplet repeats in the fragile X mental retardation gene 1 promoter.
    J Biol Chem. 1996 Oct 4;271(40):24325-8 PMID: 8798682
  71. Absence of disease phenotype and intergenerational stability of the CAG repeat in transgenic mice expressing the human Huntington disease transcript.
    Hum Mol Genet. 1996 Feb;5(2):177-85 PMID: 8824873
  72. Cryptic and polar variation of the fragile X repeat could result in predisposing normal alleles.
    Cell. 1994 Jun 17;77(6):853-61 PMID: 7911740
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.19.8.5675
Published
1999-08-00
Pages
5675-84
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC84419
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1128/MCB.19.8.5675 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