October 24, BIOS 95: Cell Division and Chromosome Dynamics
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1 October 24, 2008 BIOS 95: Cell Division and Chromosome Dynamics
2 Cancer unregulated cell growth and migration Aneuploidy errors in chromosome segregation and genome maintenance living with an altered genome
3 Aneuploidy can be limited
4 Aneuploidy can Rampant Duplications Translocations
5 Cancer unregulated growth and movement Aneuploidy errors in chromosome segregation and genome maintenance Metastasis cell migration and ignoring cues leaving home without a forwarding address
6 Shape and proximity Love the neighbor Not so close
7 Wildtype cells Tumor cells Scratch test Time Tumor cells Show Increased mobility Zhu et al 2004
8 Chromosome Segregation and Cell Migration A. Chromosome segregation - Main points: 1.The essential blue-print of instructions are contained on chromosomes (you have 2 sets of 23 and you need them ALL) 2. Before a cell divides into two the cells must copy each chromosome to obtain to identical sets 3. Cell division is the process by which the chromosomes (and later cell volume) are separated 4. Chromosome segregation requires structural elements, mechanical forces and geometry
9 Phases in the life of a cell and in making another cell Chromosome segregation
10 Chromosome segregation M G2 G1 S DNA Replication
11 Simple scopes - mitosis
12 Polarization scope Inoue et al 1995
13 Fibers required to move chromosomes Inoue et al 1995
14 Cytoskeleton: actin - cell surface shape, cell migration microtubules - internal organelle trafficking, chromosome segregation intermediate filaments - mechanical strength, resist shear
15 The soluble subunit Tubulin (that makes up Microtubules) is really two proteins tightly bound together ALL OF THE TIME Tubulin = heterodimer PEANUT
16 Tubulin heterodimers associate head-to-tail (like actin) but aggregate as 13 protofilaments into a hollow cylinder!
17 The 25 nm cylinder is strong - resists bending and compression Tubulin subunits are distinct β α because of head-to-tail assembly microtubule also has polarity β subunit end α subunit end
18 Addition promotes hydrolysis of underlying tubulin
19 Why microtubules Are not like Rock candy crystals At steady state (no net change in polymer) EACH microtubule randomly switches between growth and shortening -> Dynamic Instability
20 Microtubule plus-ends exhibit Dynamic Instability Random loss/gain of tubulin GTP cap
21 Biochemistry Identifying Mt-associated proteins
22
23 Microtubule tip (plus-end) binding proteins EB1 and Green Fluorescence Protein
24 Microtubule ends are dynamic lattice relatively immobile
25 Microtubules exhibit different rates of catastrophe and rescue throughout the cell cycle
26 Review #1 Microtubules are hollow cylinders made up of tubulin subunits Microtubule ends are biochemically different Microtubules exhibit Dynamic Instability the plus-end (β-tubulin end) randomly switches between growth and shortening Cells regulate microtubule dynamics throughout cell cycle
27 Microtubules are the railway of the cell squid axoplasm...
28
29 Microtubules generate railway system Railway is polarized Melanocyte in + Melanocyte out
30 Microtubules generate railway system Railway is polarized... Melanocyte in + Melanocyte out
31 Polarized railway... You go to the plus-end, and I ll go to the minus-end: Opposing motors
32 Microtubules are nucleated AND organized by the centrosome Notice polarized array
33 Microtubules exhibit dynamic instability plus-ends randomly switching between growth and shortening minus-ends are tethered to a pole foci Nucleation - single site
34
35 Review #2 Microtubules generate a railway system in the cell Microtubules are nucleated from centrosomes Centrosomes seed microtubule growth so that a polarized array is formed β-end (plus-end) points to cell periphery Motors transport cargo along microtubules Kinesins are dedicated plus-end directed motors Dyneins are dedicated minus-end directed motors
36 Cell division Re-organization of the microtubule railway into two polar arrays oppositely oriented Chromosomes capture microtubule ends
37
38 Pericentriolar material (not centrioles) nucleates Microtubules requires a specialized tubulin seed
39 During S-phase, centrosomes duplicate Two sets of polarized Microtubules arrays
40 Two sets of polarized Microtubules arrays Kinesins help Drive poles apart + + Anti-parallel arrays and kinesin complexes
41 Just prior to mitosis entry duplicated chromosomes condense plate of spaghetti model
42 Prophase -> Nuclear Envelope Breakdown -> Prometaphase Cytoplasmic microtubules can now be captured by chromosomes
43
44 Microtubules captured by discrete protein complexes on each Sister chromatids Skibbens et al
45 Centrosomes Microtubules Pulling Forces Kinetochores - protein complex assembled onto centromere DNA Microtubule capturing complex
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