01/27/16

MCB 104
[missed 5 min, intro]
Key features of cell division
Chromosome condensation
o Can be pulled apart and separated in equal matter
Mitotic spindle assembly
o Bipolar mt organization
o Correct chromosome attachments to kinetochores
Separation of chromosomes to daughter cells
o Putting motors and dragging things to appropriate places
Contractile ring assembly (for cytokinesis)
o Dividing the cell
Different cytoskeletal systems carry out mitosis and cytokinesis
Microtubules mitosis
Actin cytokinesis
o Actin and myosin filaments of the contractile ring
Mitosis: emphasis on chromosomes
Carry genetic information
Mitosis highlight 1: chromosome condensation
Interphase dna is diffuse in the nucleus, in extended chromatin
o Dna is assembled in histones nucleosomes
o Need to be packaged in higher order system
o During mitosis, the condensing complex drives folding of chromatin to a specific
structure
Make rings and organizes chromatin in specific ways. Stacks on top of
another
Extended structure compressed
o The packaging allows reliable segregation within the cell (necessary for
separation)
Mitotic chromosome organization
Sister chromatids are tethered together after duplication
o Lose association
Cohesion rings
o Cohesion rings keep sister chromatids tethered until anaphase, when separation is
initiated by the destruction of cohesion
o Protein complex
Centromere
o Chromosomal region where the kinetochore assembles
Kinetochore
o Junction complex between MTs and chromosome

o Near centromere
o Multiprotein complex
o Allows chromatid to attach to mitotic spindle
Nuclear envelope breakdown and chromosome attachment to spindle occur in prometaphase
Break down nuclear envelope
Microtubules can now associate with chromosomes through kinetochore
Sister chromatids align during metaphase
Activity of microtubules
Mitosis highlight 2: construction of the mitotic spindle
MTs are 10x more dynamic in mitosis than interphase\
Duplication of centrosomes
Catastrophe frequency increases
o Microtubules assemble
Average lifetime of MTs goes from 10 to 30
Microtuble organization in the mitotic spindle
3 types of MT construct a bipolar antiparallel array
astral mt
o radiate out of MTOC
o contact plasma membrane
o keepa vision axis along the plane
polar microtubules
o associate with other microtubules from other centrosome
o help position microtubule
o antiparallel manner
o position mitotic spindle
o zone of interdigitation
o prevent crashing
kinetochore mt
o attach to chromosome
o provide physical force and tracts that put tension that separate the chromosomes
Pathways of chromosome capture
search and capture
o microtubules (dynamic instability)
o mts grow and shrink, stabilized when they contact kinetochore
r heald
o take fluorescent beads. Coat in DNA
o put in cell extract with tubulin
o even in absence of real kinetochore or centrosome, DNA alone is sufficient to
organize mitotic spindle
self organization
o mts grow around chromosomes
o motors then organize them
o tetrameric kinesin-5 cross links and stabilizes MTs

 antiparallel
o dynein focuses spindle poles
focus on parallel
walking down end will focus end together
mitosis: emphasis on kinetochores
metaphase MT dynamics
dynamic stability
even when spindle appears stable:
o mt grow and shrink; chromosomes remain attached and oscillate
tension created
o poleward mt flux )treadmilling)
o these maintain tension on kinetochores
fluoresencently label express GFP tubulin
o laser > FRAP (fluorescence recovery after photobleaching)
laser kills fluorescence
allows us to make feduiciary mark on mitotic spindle
watch how blank zone changes over time
bleached zones move poleward
polymerization end and depolarization + end
spindle assembly checkpoint
o check whether theres a physical force
mitosis highlight 3: anaphase separation of chromosomes
once spindle forms, problem is to move the chromosomes to different poles
lack chromosomes
o cell dies
more chromosomes
o down syndrome, cancer
initiating anaphase
apc activation, cohesin cleavage and chromosome separation
spindle aseembly checkpkoint. Read by tension on KT
o balance of forces
if sister KTs are not attached to opposite spindle poles delay in metaphase
if sister KTs are attached to opposite spindle poles go to anaphase activates APC
apc- uniquitylates securin degraded
o anaphase promoting complex
separase protease active cleaves cohesion sister chromatids separate
mt dynamics during anaphase
shortening of kinetochore mts pulls chromosomes apart
o pull chromatids toward poles
o stop + end addition
elongation of interpolar mts separates poles
o pushed two poles apart from one another
anaphase chromosome movements

 mt depoalymerization of kinetochore but it stays ttached to shrinking mt pacman

mitosis highlight 4: cytokinesis
strong ring of actin that forms former mitotic plate
recruits myosin and pinches off membrane
regulated by rho protein