Analysis of the cell cycle in the budding yeast Candida albicans by positioning of chromosomes by fluorescence in situ hybridization (FISH) with repetitive sequences

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Abstract
In the budding yeasts, including Saccharomyces cerevisiae, in which individual chromosomes cannot be visualized by microscopy, the mitotic phases in the cell cycle have not been correlated with the chromosome behaviour. We used various repetitive sequences, namely, rDNA, telomeric sequences and RPSs, which are localized in limited regions in almost all chromosomes, as probes for fluorescence in situ hybridization (FISH) to analyse the cell cycle phases in a pathogenic yeast Candida albicans. The positioning of the FISH signals was analysed quantitatively in relation to the length of spindle microtubules in the nuclear domain.RPSs were randomly distributed in the interphase nucleus, and they formed aggregates with the development of the spindle. DNA synthesis was complete before RPSs came closest to the spindle. As the spindle elongated, they were scattered along the spindle and then separated into two clusters at the spindle poles at the end of anaphase. rDNA was localized in the nucleolar domain, and telomere signals were randomly distributed throughout mitosis.By estimating quantitatively the proportions of mitotic cells with particular configurations of both microtubules and chromosomes in a population of rapidly proliferating cells, we were able to define various stages in the progression of mitosis. The S phase and pro-to-prometaphase were overlapping and the G2 phase was lacking. Unexpectedly, the pole-to-pole elongation of the spindle (anaphase B) was predominating and was followed by movement of chromosomes to the poles (anaphase A).
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