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

2. YEAST EM

4. CELL POLARITY STIMULUS RESPONSE CONTENT DISTRIBUTION FUNCTIONAL DOMAIN

5. BUD6-GFP

6. 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

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

8. ROBOTIC MICROSCOPY

9. DHC-GFP Sid Shaw 1997 NUCLEAR DIVISION

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

12. ASH1 mRNA LOCALIZATION

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

14. RNA LABELING SYSTEM

15. gRNA ASH1 LOCALIZATION

17. bni1  gRNA

18. bud6  gRNA

19. WTmyo4/she1bni1/she5bud6/aip3 TIME COMPOSITE IMAGES

20. CORTICAL SCAFFOLD

21. Kar9-GFPBud6-GFPASH1 mRNA MT orientation during vegetative growth and mating Bud site selection and actin filament orientation Daughter specific inhibition of HO transcription Polarized Determinants

22. THANKS !!!