Abstract |
Aneuploidy and gross chromosomal rearrangements (GCRs) can lead to genetic diseases and the development of cancer. We previously demonstrated that introduction of the repetitive retrotransposon Ty912 onto a nonessential chromosome arm of Saccharomyces cerevisiae led to increased genome instability predominantly due to increased rates of formation of monocentric nonreciprocal translocations. In this study, we adapted Multiplex Ligation-dependent Probe Amplification (MLPA) to analyze a large numbers of these GCRs. Using MLPA, we found that the distribution of translocations induced by the presence of Ty912 in a wild-type strain was nonrandom and that the majority of these translocations were mediated by only six translocation targets on four different chromosomes, even though there were 254 potential Ty-related translocation targets in the S. cerevisiae genome. While the majority of Ty912-mediated translocations resulted from RAD52-dependent recombination, we observed a number of nonreciprocal translocations mediated by RAD52-independent recombination between Ty1 elements. The formation of these RAD52-independent translocations did not require the Rad51 or Rad59 homologous pairing proteins or the Rad1-Rad10 endonuclease complex that processes branched DNAs during recombination. Finally, we found that defects in ASF1-RTT109-dependent acetylation of histone H3 lysine residue 56 (H3K56) resulted in increased accumulation of both GCRs and whole- chromosome duplications, and resulted in aneuploidy that tended to occur simultaneously with GCRs. Overall, we found that MLPA is a versatile technique for the rapid analysis of GCRs and can facilitate the genetic analysis of the pathways that prevent and promote GCRs and aneuploidy.
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Authors | Jason E Chan, Richard D Kolodner |
Journal | PLoS genetics
(PLoS Genet)
Vol. 8
Issue 3
Pg. e1002539
( 2012)
ISSN: 1553-7404 [Electronic] United States |
PMID | 22396658
(Publication Type: Journal Article, Research Support, N.I.H., Extramural)
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Chemical References |
- ASF1 protein, S cerevisiae
- Cell Cycle Proteins
- Histones
- Molecular Chaperones
- RAD52 protein, S cerevisiae
- Rad52 DNA Repair and Recombination Protein
- Retroelements
- Saccharomyces cerevisiae Proteins
- Histone Acetyltransferases
- Rtt109 protein, S cerevisiae
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Topics |
- Acetylation
- Aneuploidy
- Base Sequence
- Cell Cycle Proteins
(genetics, metabolism)
- Chromosome Duplication
(genetics)
- Genome, Fungal
- Genomic Instability
- Histone Acetyltransferases
(genetics, metabolism)
- Histones
(genetics)
- Molecular Chaperones
(genetics, metabolism)
- Molecular Sequence Data
- Multiplex Polymerase Chain Reaction
(methods)
- Rad52 DNA Repair and Recombination Protein
(genetics, metabolism)
- Recombination, Genetic
- Retroelements
- Saccharomyces cerevisiae
(genetics)
- Saccharomyces cerevisiae Proteins
(genetics, metabolism)
- Translocation, Genetic
(genetics)
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