Duplication processes in Saccharomyces cerevisiae haploid strains - PubMed (original) (raw)

Duplication processes in Saccharomyces cerevisiae haploid strains

Joseph Schacherer et al. Nucleic Acids Res. 2005.

Abstract

Duplication is thought to be one of the main processes providing a substrate on which the effects of evolution are visible. The mechanisms underlying this chromosomal rearrangement were investigated here in the yeast Saccharomyces cerevisiae. Spontaneous revertants containing a duplication event were selected and analyzed. In addition to the single gene duplication described in a previous study, we demonstrated here that direct tandem duplicated regions ranging from 5 to 90 kb in size can also occur spontaneously. To further investigate the mechanisms in the duplication events, we examined whether homologous recombination contributes to these processes. The results obtained show that the mechanisms involved in segmental duplication are RAD52-independent, contrary to those involved in single gene duplication. Moreover, this study shows that the duplication of a given gene can occur in S.cerevisiae haploid strains via at least two ways: single gene or segmental duplication.

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Figures

Figure 1

Figure 1

Correspondence between the simplified restriction map of the URA2 locus and protein domains. The thick line indicates the the URA2 coding sequence (restriction site: b, BglII; B, BamHI). A, C, D, B and E denote the BamHI–BglII subfragments, ns15 and ns30 the positions of the nonsense mutations, and fs72 the position of the frameshift mutation (GATase, glutamine amidotransferase; CPSase, carbamoylphosphate synthetase; DHOase-like, dihydroorotase-like; ATCase, aspartate transcarbamylase).

Figure 2

Figure 2

Chromosome location of the duplication events. (A) PFGE of the intact chromosomes from the ancestral strain and each of the revertant strains. (B) Hybridization of the corresponding Southern blot with the ATCase probe.

Figure 3

Figure 3

Size of the duplicated region. Diagrams give the profiles obtained upon preforming CGH on to microarray. The _x_-axis correspond to ORFs from the left arm of the chromosome X running from YJL001w to YJL204c. The _y_-axis correspond to the genomic ratio calculated between the revertant and ura2 15-30-72 strain.

Figure 4

Figure 4

Chromosomal insertion of the segmental duplicated regions. The grey boxes correspond to the duplicated elements whereas the dotted boxes correspond to the original region. The black arrow indicates the duplicated part of the URA2 gene. The numbers placed above the duplicated structure correspond to the size of the duplicated segment.

Figure 5

Figure 5

Sequence of the 5′ junction. The numbers placed above and below the sequence correspond to the chromosomal coordinates. The bold face type indicates a short region of identity.

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