Getting it right: How your mitotic cells divide your chromosomes accurately. Dr. Cassimeris Biological Science
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1 Getting it right: How your mitotic cells divide your chromosomes accurately Dr. Cassimeris Biological Science
2 Cell division is a normal function that maintains our tissues and organs Bone marrow stem cells >1,000,000 divisions/min Skin stem cells Intestinal stem cells Muscle satellite cells Liver cells
3 Why do cells need to accurately copy and divide DNA at each cell division? Weaver and Cleveland. Cancer Research
4 Excessive genetic instability is deadly Weaver and Cleveland. Cancer Research
5 Cells multiply by dividing the cell division cycle One cell in mitosis
6 Mitosis Overview prometaphase metaphase anaphase
7 Some of the most successful chemotherapies target mitosis Taxol Vinblastine
8 Goals for today: How do cells divide chromosomes? How do cells monitor chromosomes to know that each daughter cell will get one copy of the genome? How do chemotherapies block cell division? Why does a block in cell division lead to cell death?
9 How can a cell pair each chromosome with its exact copy? How can a cell know that all chromosomes are attached to the mitotic spindle before they divide?
10 How can a cell pair each of its chromosomes with the exact copy? Answer: Attach each chromosome to its copy as the copy is made. This happens in S-phase
11 Cohesins: the glue that holds chromosomes chromosome together cohesins only let go at the right time Cohesins must hold on for ~ 12 hrs Cohesin proteins Cohesin proteins hold chromosomes together starting here
12 How can a cell pair each chromosome with its exact copy? How can a cell know that all chromosomes are attached to the mitotic spindle before they divide?
13 breaking cohesions and moving chromosomes to opposite poles only happens when ALL chromosomes are properly attached to the spindle
14 To understand how the system functions, we need a few critical parts Pole or centrosome Kinetochore microtubule
15 A real human cell Pole Microtubule Chromosome Kinetochore
16 How do chromosomes make correct attachments when mitosis begins?
17 Microtubules: The Cell s Bones a simple polymer MTs, DNA, Pole
18 What Microtubules Do- Find Things Move Things
19 By growing and shortening, microtubules find kinetochores on chromosomes by a random search & capture process
20 summary Form attachments Check attachments Break cohesions & move to opposite poles
21 Metaphase to Anaphase Transition Microtubules depolymerize Cohesions destroyed & motors pull chromosomes to poles Metaphase Anaphase motor protein
22 Microtubules depolymerize Cohesions destroyed & motors pull chromosomes to poles Metaphase Anaphase This transition step happens for all chromosomes simultaneously, but only when ALL chromosomes (46 in our cells) are attached to the spindle correctly
23 How could a cell know that ALL chromosomes are ready to divide? one example of a cell cycle checkpoint WAIT checkpoints turn on wait signals
24 Kinetochores are the detectors of chromosome attachment to the spindle kinetochore
25 How do we know that kinetochores are the site of detection? SAC = spindle assembly checkpoint What would happen if we destroyed the unattached kinetochore?
26 How do we know that kinetochores are the site of detection? SAC = spindle assembly checkpoint What would happen if we destroyed the unattached kinetochore? Cell immediately enters anaphase
27 Kinetochores sense stable microtubule attachments and tension pulling forces stretch chromatin & stabilize microtubule attachments the distance between kinetochores is a proxy for tension
28 WAIT signal on until all chromosomes attached properly Correct attachments. WAIT signal removed. Metaphase (Wait signal ON) Anaphase (Wait signal OFF)
29 So, how do chemotherapies that target mitosis work to kill cancer cells? Taxol Vinblastine
30 Taxol & other microtubule-targeted drugs block microtubule turnover & prevent proper microtubule-kinetochore attachments normal cell Pure tubulin Taxol-treated cells + tubulin-binding chemotherapy Vasquez et al MBoC. 8: 973. Basically these drugs ACTIVATE the SAC
31 A cell that stays in mitosis too long activates apoptosis prolonged mitotic block Tb DNA Wait signal stays ON in taxol
32 Timing is everything. Cells will stall in mitosis to fix chromosome attachments. But if this process takes too long, cell death results. Normal Mitosis Metaphase CDK1/cyclinB Anaphase X CDK1/cyclinB CDK1 active for short time, apoptosis blocked Prolonged Mitosis CDK1/cyclinB Caspases CDK1 active for long time activates caspase enzymes (apoptosis)
33 want to learn more while at Lehigh? BioS 41: Introduction to cell, molecular biology and biochemistry. BioS 367: Cell Biology Advanced courses by Drs. Skibbens or Cassimeris
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