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M267 Lecture 2, Cell Cycle February 27, 2006

M267 Lecture 2, Cell Cycle February 27, 2006. Chicken cells. Human cells. Flu virus or polyethylene glycol. S phase cells. G 1 phase cells. 3 H-TdR. Mitotic cell. G1 , S , or G2 interphase cell. Premature Chromosome Condensation. C. A3. A2. A1. B. Fraction number.

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M267 Lecture 2, Cell Cycle February 27, 2006

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  1. M267 Lecture 2, Cell Cycle February 27, 2006

  2. Chicken cells Human cells Flu virus or polyethylene glycol

  3. S phase cells G1 phase cells 3H-TdR

  4. Mitotic cell G1, S, or G2 interphase cell Premature Chromosome Condensation

  5. C A3 A2 A1 B Fraction number tryptophan arginine lysine

  6. Hela cells were sychronized at M/G1 (you know how to do this) 3 Rate of DNA synthesis 2 1 M/G1

  7. Fraction number

  8. G1 + S + G2 G1 + S + G2 G1 + S + G2 M M M

  9. The size of the bud identifies where the cells are in progress through the cell cycle

  10. Shift to non-permissive temperature G1 M G2 S Mutant 1

  11. Shift to non-permissive temperature G1 M G2 S Mutant 2

  12. Shift to non-permissive temperature G1 M G2 S “Execution Point” Mutant 3

  13. Shift to non-permissive temperature G1 M G2 S This is a cdc mutant that arrests at G2/M Mutant 4

  14. cdc28 arrest START M Cerevisiae

  15. A population of S. pombe cdc2 mutant cells after shifting to the non-permissive temperature for a period greater than one cell cycle.

  16. M M

  17. “Expression” cDNA library in which human protein coding sequences can be expressed in yeast cells Only 5 cells out of 106 form colonies Permissive temperature Non-permissive temperature

  18. G1 S G2 Percent cells in each phase 2 4 6 8 10 12 Fraction p34 (cdc2 kinase) p62 p34 p62 casein

  19. + Cdc2 cyclin cyclin Cdc2 p34 p34 Inactive kinase active cyclin-dependent kinase

  20. Cdc2 cyclin p34 HISTONE + ATP HISTONE-P + ADP

  21. Cyclin Histone 2A 0 2 4 6 8 10 12 14 16 18 20 22 24 G2/M peak Hours after release from G1/S Hela cells were sychronized at G1/S (you know how to do this).

  22. Cyclin 0 2 4 8 16 Hours after release from mitosis Hela cells were sychronized at M/G1 (you know how to do this).

  23. Cyclin protein 0 2 4 6 8 10 12 14 16 18 20 22 24 G2/M peak Hours after release from G1/S Intensity of cyclin, abitrary units 10 15 20 30 Hours after release from G1/S Hela cells were sychronized at G1/S

  24. Cyclin protein 0 0.5 1.0 1.5 2 4 5 Hours after release from mitosis Intensity of cyclin, abitrary units 0 0.5 1.0 1.5 2 4 5 Hours after release from mitosis Hela cells were sychronized at M/G1

  25. Cyclin A Cyclin B

  26. Cdc2 Cdc2 Cdc2 Substrate g Substrate h Substrate i Substrate a Substrate b Substrate c Substrate d Substrate e Substrate f

  27. Cyclin??? M G1 Cyclin A&B S G2 Cyclin Cyclin B B cdc2 CDK? cdc2 cdc2 cdc2 p p

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