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DYNAMICS OF POLARIZED DETERMINANTS AND NUCLEAR SEGREGATION IN YEAST

DYNAMICS OF POLARIZED DETERMINANTS AND NUCLEAR SEGREGATION IN YEAST. (Or Yeast “Pong”). Dale Beach Kerry Bloom Lab. March 4, 2000. YEAST EM. CELL POLARITY. STIMULUS RESPONSE CONTENT DISTRIBUTION FUNCTIONAL DOMAIN. BUD6-GFP. NUCLEAR DIVISION. Pushing forces in unbudded cells

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DYNAMICS OF POLARIZED DETERMINANTS AND NUCLEAR SEGREGATION IN YEAST

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  1. DYNAMICS OF POLARIZED DETERMINANTS AND NUCLEAR SEGREGATION IN YEAST (Or Yeast “Pong”) Dale Beach Kerry Bloom Lab March 4, 2000

  2. YEAST EM

  3. CELL POLARITY • STIMULUS RESPONSE • CONTENT DISTRIBUTION • FUNCTIONAL DOMAIN

  4. BUD6-GFP

  5. NUCLEAR DIVISION • Pushing forces in unbudded cells • Locate the bud • Penetrate the neck • Pulling forces towards the bud • Oscillations at the neck • Spindle Elongation • Spindle Disassembly

  6. LIVE CELL ANALYSIS • GFP: in-vivo Protein Labeling • Follow Sequential Processes • Record Dynamics

  7. ROBOTIC MICROSCOPY

  8. NUCLEAR DIVISION DHC-GFP Sid Shaw 1997

  9. NUCLEAR DIVISION

  10. NUCLEAR MIGRATION DEFECT • Bud Specific • KAR9 • BUD6/AIP3 • BNI1/SHE5 • Mother Specific • NUM1 • Unknown Localization • DHC1/DYN1

  11. ASH1 mRNA LOCALIZATION

  12. CODING 3’ UTR PRO ASH1 LOCALIZATION ELEMENTS ASH1 mRNA mRNA “Zip Codes” Chartrand et al., 1998; Gonzalez et al., 1998

  13. RNA LABELING SYSTEM

  14. gRNAASH1 LOCALIZATION

  15. gRNAASH1 LOCALIZATION

  16. bni1D gRNA

  17. bud6D gRNA

  18. WT myo4/she1 bni1/she5 bud6/aip3 TIME COMPOSITE IMAGES

  19. CORTICAL SCAFFOLD

  20. Polarized Determinants Kar9-GFP Bud6-GFP ASH1 mRNA MT orientation during vegetative growth and mating Bud site selection and actin filament orientation Daughter specific inhibition of HO transcription

  21. THANKS !!!

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