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Saccharomyces cerevisiae Budding Yeast

Saccharomyces cerevisiae Budding Yeast. Movie. 4-5 um. minutes. Mitochondria. EM view. Bud site. Nucleolus. ER/ Golgi. Nucleus. Vacuole. Nuclear envelope And Nuclear Pores. Wall. ~5 um. The Yeast Nuclear Genome. bp. 16 Chromosomes (200-1600 Kbp)

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Saccharomyces cerevisiae Budding Yeast

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  1. Saccharomyces cerevisiaeBudding Yeast Movie 4-5 um minutes

  2. Mitochondria EM view Bud site Nucleolus ER/ Golgi Nucleus Vacuole Nuclear envelope And Nuclear Pores Wall ~5 um

  3. The Yeast Nuclear Genome bp 16 Chromosomes (200-1600 Kbp) 12,052 Kbp nuclear genome (3.5x E. coli) roughly 6000 protein-encoding genes (the mitochondrial genome is about 76,000bp and encodes about 50 ORFs)

  4. Transcriptome Profiling

  5. Human Disease Homologs Yeast TBLASTN Percent Percent Human Gene ID Gene P-value Identity Similarity MSH2 3.8e-255 43 65 Mutator gene (MSH2, colon cancer) YCF1 2.4e-157 31 57 Cystic fibrosis conductance regulator (CFTR) GEF1 3.4e-95 33 58 Voltage-gated chloride ion channel TEL1 8.8e-84 49 36 Ataxia telangiectasia gene YNL161W 8.5e-82 41 65 Myotonic dystrophy associated protein kinase SOD1 8.9e-56 55 69 Superoxide dismutase (SOD-1) SGS1 3.1e-50 24 34 Werner's Syndrome gene IRA2 1.0e-28 21 45 Neurofibromin (NF1)

  6. The Domestication of Yeast

  7. The Yeast Life Cycle “sex and the single yeast” Diploid (2n)Haploid (n) Alternation of Generations Mating (Conjugation) heterothallic mother cell in the mother cell

  8. The Mitotic Cell Cycle

  9. Master Regulatory Molecules Control the Cycle Transcription Cyclins (9 total) + CDK (Cdc28) factors 90 minute generation time

  10. Controlling S Ori Licensing Start

  11. Bud Site Selection by Cdc42

  12. A Cdc42ts Mutant is Defective in Budding and Division Superposition of Spc42::GFP to mark MTOCs and Septin:GFP to mark bud site

  13. Septins Ring the Bud Site Septin::GFP

  14. MicrofilamentsTarget the Bud Cortical Patches Target Exocytosis Contractile Ring at Cytokinesis Long Cables Support Transport

  15. The Secretory Pathway Delivers

  16. Localized Wall Synthesis Controls Growth Buds Movie

  17. An Intra-Nuclear Spindle

  18. Spc42::GFP DNA The Spindle Pole Body The Fungal MTOC

  19. The Cytoskeleton in Mitosis Tubulin::GFP Myosin1(MyosinII)::GFP

  20. homotypic mutants can’t switch Mothers Switch in G1 Mating Type Switching After Cytokinesis A A HO Alpha A HO HO A Alpha Alpha A

  21. alpha a a Silent Active Silent HML MAT HMR Switching is a Gene Conversion Event alpha alpha a alpha a a Recombination Enhancer Determines the Polarity of the Switch Silenced Silenced Switching Switching at MAT Changes the Mating Type of the Cell

  22. Certain mRNAs and proteins are Transported into the Bud Myosin V Ash1 protein Ash1 mRNA encodes a transcriptional repressor of HO expression, suppressing switching in the daughter cell HO endonuclease

  23. Mating also involves polarized growth Movie

  24. Pheromone response Cdc42 MAPKKKK MAPKKKMAPKKMAPK Ste5 is a scaffolding protein responsible for organizing a MAP kinase cascade CDC42 G-protein FUS proteins

  25. Hyphal Growth(Invasive) Candida albicans

  26. A Few Questions for Thought • Describe the life cycle of Saccharomyces cerevisiae, noting relationships between mitotic growth, the alternation of generations, meiosis, conjugation and spore formation. • Review the compartments and cytoskeleton of the eukaryotic cell, comparing and contrasting what you learned in the first semester with this model yeast. • Compare and contrast (in a simple way) mammalian and fungal cell cycles. • Describe the role(s) of septins, spindle pole bodies, and CDC42 protein in the growth of E. coli. • How is a cell’s mating type identity determined? How can it switch (and why)? How is switching repressed in certain cells? Why is this important? • (After lab) Explain how a cell senses the presence of a mating partner, and what happens once it shows interest?

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