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Molecular and Cellular Biology, August 1999, p. 5675-5684, Vol. 19, No. 8
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Stability of the Human Fragile X (CGG)n Triplet Repeat Array in Saccharomyces cerevisiae Deficient in Aspects of DNA Metabolism

Peter J. White,1,dagger Rhona H. Borts,2,Dagger and Mark C. Hirst1,*

Fragile X Group1 and Yeast Genetics Laboratory,2 Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Headington, Oxford OX3 9DS, United Kingdom

Received 22 September 1998/Returned for modification 18 November 1998/Accepted 11 May 1999

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.

* Corresponding author. Mailing address: Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Headington, Oxford OX3 9DS, United Kingdom. Phone: 44-1865-222437. Fax: 44-1865-222500. E-mail: mhirst{at}worf.molbiol.ox.ac.uk.

dagger Present address: Department of Biological & Molecular Sciences, University of Stirling, Stirling FK9 4LA, United Kingdom.

Dagger Present address: Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom.

Molecular and Cellular Biology, August 1999, p. 5675-5684, Vol. 19, No. 8
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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