A Hayashi1, H Ogawa, K Kohno, S M Gasser, Y Hiraoka. 1. Kansai Advanced Research Center, Communications Research Laboratory, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe, Japan.
Abstract
BACKGROUND: Meiosis is a process of universal importance in eukaryotic organisms, generating variation in the heritable haploid genome by recombination and re-assortment of chromosomes. The intranuclear movement of chromosomes is expected to achieve pairing and recombination of homologous chromosomes during meiosis. Meiosis in the budding yeast Saccharomyces cerevisiae has been extensively studied, both genetically and by molecular biology; here we report cytological observations of meiotic chromosomal events in this organism. RESULTS: Using fluorescence microscopy, we have examined the behaviour of chromosomes and microtubules during meiosis in S. cerevisiae. We first observed the dynamic behaviour of nuclei in living cells using jellyfish green fluorescent protein (GFP) fused with nucleoplasmin, a Xenopus oocyte nuclear protein. The characterization of nuclear movement in living cells was extended by an analysis of chromosomes and microtubules in fixed specimens. In addition, the nuclear localization of centromeres and telomeres was determined by indirect immunofluorescence microscopy in synchronous populations of meiotic cells. While telomeres remain in clusters of 5-8 throughout meiosis, centromeres change their nuclear localization dramatically during the progression of meiosis: centromeres are first clustered at a single site near the spindle-pole body before the induction of meiosis, and become scattered during the meiotic prophase. CONCLUSIONS: Our observations have demonstrated that nuclear and cytoskeletal reorganization take place with meiosis in S. cerevisiae. In particular, the distinct relocalization of centromeres during meiosis indicates a considerable movement of chromosomes within the meiotic prophase nucleus.
BACKGROUND: Meiosis is a process of universal importance in eukaryotic organisms, generating variation in the heritable haploid genome by recombination and re-assortment of chromosomes. The intranuclear movement of chromosomes is expected to achieve pairing and recombination of homologous chromosomes during meiosis. Meiosis in the budding yeastSaccharomyces cerevisiae has been extensively studied, both genetically and by molecular biology; here we report cytological observations of meiotic chromosomal events in this organism. RESULTS: Using fluorescence microscopy, we have examined the behaviour of chromosomes and microtubules during meiosis in S. cerevisiae. We first observed the dynamic behaviour of nuclei in living cells using jellyfish green fluorescent protein (GFP) fused with nucleoplasmin, a Xenopus oocyte nuclear protein. The characterization of nuclear movement in living cells was extended by an analysis of chromosomes and microtubules in fixed specimens. In addition, the nuclear localization of centromeres and telomeres was determined by indirect immunofluorescence microscopy in synchronous populations of meiotic cells. While telomeres remain in clusters of 5-8 throughout meiosis, centromeres change their nuclear localization dramatically during the progression of meiosis: centromeres are first clustered at a single site near the spindle-pole body before the induction of meiosis, and become scattered during the meiotic prophase. CONCLUSIONS: Our observations have demonstrated that nuclear and cytoskeletal reorganization take place with meiosis in S. cerevisiae. In particular, the distinct relocalization of centromeres during meiosis indicates a considerable movement of chromosomes within the meiotic prophase nucleus.