Unknown,Transcriptomics,Genomics,Proteomics

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Genotyping by array of yeast to map post-meiotic segregation across the yeast genome

ABSTRACT: To map post-meiotic segregation (PMS) across the yeast genome, we genotyped the two cells resulting from the first mitotic division of the four spores of 4 tetrads of a YJM789/S96 Saccharomyces cerevisiae hybrid strain. Sporulation was induced, tetrads were dissected, spores let to germinate and the two cells coming from the first mitotic division of each spore were finally dissected. DNA from each of the eight cells in each tetrad was extracted from independent overnight cultures in rich medium and hybridized to microarrays, one array per cell. Each hybridization was used to genotype the corresponding cell and genetic differences between the two cells from the same spore revealed PMS. Therefore there are 32 hybridization files, 2 per spore and 8 per tetrad.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Charles Girardot

PROVIDER: E-TABM-1031 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Spermatophyte pollen tubes and root hairs have been used as single-cell-type model systems toward understanding the molecular processes underlying polar growth of plant cells. Horsetail (Equisetum arvense L.) is a perennial herb species in Equisetopsida, which creates separately growing spring and summer stems in its life cycle. The mature chlorophyllous spores produced from spring stems can germinate without dormancy. Here we report the cellular features and protein expression patterns in five stages of horsetail spore germination (mature spores, rehydrated spores, double-celled spores, germinated spores, and spores with protonemal cells). Using 2-DE combined with mass spectrometry, 80 proteins were found to be differentially expressed upon spore germination. Among them, proteins involved in photosynthesis, protein turnover, and energy supply were over-represented. Thirteen proteins appeared as isoforms on the gels, indicating the potential importance of post-translational modification. In addition, the dynamic changes of ascorbate peroxidase, peroxiredoxin, and dehydroascorbate reductase implied that reactive oxygen species homeostasis is critical in regulating cell division and tip-growth. The time course of germination and diverse expression patterns of proteins in photosynthesis, energy supply, lipid and amino acid metabolism indicated that heterotrophic and autotrophic metabolism were necessary in light-dependent germination of the spores. Twenty-six proteins were involved in protein synthesis, folding, and degradation, indicating that protein turnover is vital to spore germination and rhizoid tip-growth. Furthermore, the altered abundance of small G protein Ran1, 14-3-3 protein, actin, and Caffeoyl-CoA O-methyltransferase revealed that signaling transduction, vesicle trafficking, cytoskeleton dynamics, and cell wall modulation were critical to cell division and polar growth. These findings provide up-to-date evidences for understanding fern spore asymmetric division and rhizoid polar growth.

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Genotyping by array of yeast to map post-meiotic segregation across the yeast genome

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