Youngtae Jeong, M.D., Ph.D.

DGIST New Biology

3류 리더는 자기의 능력을 이용하고

2류 리더는 타인의 능력을 이용하고

1류 리더는 타인의 지혜를 이용한다.

• Understand the principles of stage progression and their
regulators of cell cycle

• Understand the concept and regulators of apoptosis

• S phase (DNA Synthesis) – chromosome duplication occurs
• M phase (Mitosis) – nuclear division and cytokinesis
• Prophase – sister chromatid condensation
• Prometaphase – breakdown of nuclear envelope
• Metaphase – sister chromatids are attached to opposite poles of mitotic spindle and
align at the spindle equator
• Anaphase – separate sister chromatids
• Telophase – spindle is disassembled and the segregated chromosomes are
packaged into separate nuclei
• Gap phases allow time for cell growth and to monitor internal and external conditions
• G1 – between M and S phases
• G2 – between S and M phases
• G1, S, G2 = interphases

• G0 phase – resting state from G1 phase

Q) How to tell what stage a cell has reached in the cell cycle?

A) 1. Look at the microscope

2. Staining cell with DNA-binding fluorescent dyes or antibodies

- BrdU

3. Measuring DNA concent

• The cell-cycle control system operates much like a timer or switch that triggers the
events of the cell cycle in a set sequence
1. The switches are generally binary (on/off) and launch events in a complete,
irreversible fashion
2. The cell-cycle control system is very robust and reliable, partly because backup
mechanisms allow the system to operate effectively
3. The control system is highly adaptable and can be modified to suit specific cell types
or to respond to specific intracellular or extracellular signals
• Cyclin – Cdk regulators, being synthesized and degraded
• Cdk – constantly expressed but active only when binds to cyclins
• Cyclins: G1-cyclins, G1/S-cyclins, S-cyclins, M-cyclins
• Cdks: G1-Cdk, G1/S-Cdk, S-Cdk, M-Cdk
• Cyclins: G1-cyclins, G1/S-cyclins, S-cyclins, M-cyclins
• Cdks: G1-Cdk, G1/S-Cdk, S-Cdk, M-Cdk
• Cdk activity regulation
1. Cyclin
2. 2nd phosphorylation by Wee1 kinase inhibits vs dephosphorylation by Cdc25
increases Cdk activity
3. Cdk inhibitor proteins (CKIs) such as p27 stimulate rearrangement of Cdk sturcture
1. Anaphase-promoting complex or cyclosome (APC/C)
- a ubiquitin ligase
- ubiquitylate and destruct securin and S- and M-cyclins
2. SCF
- another ubiquitin ligase
- ubiquitylate CKI proteins and destruct G1/S-cyclins in early S phase
• The cell cycle control system functions as a network of biochemical switches
• Cautions in DNA replication

1. Replication must occur with extreme accuracy to minimize the risk of mutations

2. Every nucleotide in the genome must be copied once

• To ensure chromosome duplication occurs only once per cell cycle
1. In early G1 phase, DNA helicases are loaded onto the replication origin and form
prereplicative complex (preRC) – DNA replication occurs only at origin
2. In S phase, DNA is unwound and the origin cannot be reused until the next G1
• Chromosome duplication requires duplication of chromatic structure but the
mechanisms are not well known.
• Cohesins hold sister chromatids together
• Mitosis and cytokinesis
• Five stages of mitosis – prophase, prometaphase, metaphase, anaphase, telophase

• Mitosis can be divided into two major parts

- abrupt increase in M-Cdk activity at G2/M transition – assembly of mitotic spindle and
its attachment to the sister-chromatids
- APC/C triggers securing destruction and leading to cohesion cleavage at meta-to-
anaphase  cyclin destruction and Cdk inactivation
• Dephosphorylation activates M-Cdk at the onset of mitosis
• To separate sister chromatids safely, two step processes occur: chromosome
condensation and sister-chromatid resolution
• Condensin regulates these processes
• Mitotic spindle is a microtubule-based machine.
• Minus ends focus at the spindle poles and plus ends radiate outward from the poles
• Kinesin-5: interacts with the plus ends of antiparallel microtubules in the spindle
midzone and push the poles apart
• Kinesin-14: minus-end directed motors, cross-link interpolar microtubules at the
spindle midzone and pull the poles
• Kinesin4 and -10: plus-end directed motors and push the attached chromosome
away from the pole
• Dyneins: minus-end directed motors and organize microtubules at various locations
in the cell
• Centrosome consists of a cloud of amorphous material and surrounds a pair of
centrioles, which are aligned longitudinally.
• Centrosome duplication begins at about the same time as the cell enters S phase
• G1/S-Cdk helps initiate centrosome duplication
• Mitotic chromosomes promote bipolar spindle assembly, using Ran-GTP
• Since chromosomes help organize the spindle microtubule, even though centrosome
is removed, the chromosome segregation occurs.

• However, without centrosome the spindle can be mispositioned

• Spindle microtubules are attached to each chromatid at kinetochore
• Ndc80 connects microtubules and kinetochore
• Kinetochore attachment
Q) How is the biorientation achieved?

A) Sister kinetochores are constructed in a back-to-back orientation

Q) How does the kinetochore sense a correct attachment of microtubules?

a) It senses a moderate level of tension

• Multiple forces act on chromosomes in the spindle
• First major force pulls the kinetochore and its associated chromatid toward the
spindle pole
• A second force is provided by microtubule flux: microtubules are pulled toward the
spindle poles and dismantled at the minus end
• A third force is the polar ejection force pushes the chromosomes away from the
spindle poles
• APC/C triggers sister chromatid separation and the completion of mitosis
• Chromosome segregate in anaphase A and B
• Anaphase A: initial poleward movement of chromosome
• Anaphase B: Separation of spindle poles
• Two sets of chromosomes are packaged into a pair of daughter nuclei

• Disassembly of mitotic spindle and reformation of nuclear envelope

• Actin and myosin II in the contractile ring generate the force for cytokinesis
• RhoA promotes actin filament formation, myosin II assembly, and ring contraction
• The microtubules of the mitotic spindle determine the plane and timing of the animal
cell division
• In the plant, cytoplasm is partitioned from inside out by a construction of a new cell
wall, called cell plate
Q) How do the various membrane-enclosed organelles segregate during cell division?

A) Mitochondria – Cells have multiple mitochondria, so they are equally distributed and
in each cycle, the number of mitochondrias doubles

B) ER – it remains intact and is cut in two during cytokinesis

C) Golgi – reorganized, fragmented and reconstructed

• Mitosis without cytokinesis – some cells undergo multiple rounds of nuclear division
without intervening cytoplasmic division

• A cell in which multiple nuclei share the same cytoplasm is called syncytium
• Later membranes are created around each nucleus in one round of coordinated
cytokinesis called cellularization

• Megakaryocytes, hepatocytes, cardiac myocytes, Drosophila embryo

• G1 phase is a stable state of Cdk inactivity
• M-Cdk inactivation is due to Cdc20-APC/C and CKI production
• Meiosis includes two rounds of chromosome segregation
• Duplicated homologs pair during meiotic prophase, forming a four-chromatid
structure called a bivalent
• Homolog pairing culminates in the formation of a synaptonemal complex
• Five divisions of meiotic prophase
1. Leptotene – homologs condense and pair, and genetic recombination begins
2. Zygotene – synaptonemal complex begins to assemble and recombination events
are occurring
3. Pachytene – the homologs are synapsed along their entire length
4. Diplotene – disassembly of the synaptonemal complexes and shortening of the
chromosomes
• In meiosis I, two sister kinetochores are fused into a single microtubule-binding unit
that attaches to just one pole
• Cleavages of cohesins along sister chromatids and at the centromeres occur at
different stages
• Organ and body size are determined by cell growth, cell division, and cell survival

• Extracellular signal molecules that regulate cell growth, division, and survival can be
categorized into three major classes

• Mitogens – stimulate cell division

• Growth factors - stimulate cell growth
• Survival factors – suppress programmed cell death
• Mitogens stimulate G1-Cdk and G1/S-Cdk activities
• DNA damage blocks cell division – The DNA damage response (DDR)
• Replicative cell senescence
– Most of human cells stop proliferation after the certain number of cell cycle
– the telomere shortening and exposed chromosome ends are sensed as DNA
damage
 activates a p53-dependent cell-cycle arrest
• Abnormal proliferation signals cause cell-cycle arrest or apoptosis, except in cancer
cells
• Proliferating cells usually coordinate their growth and division
Youngtae Jeong, M.D., Ph.D.

DGIST New Biology

3류 리더는 자기의 능력을 이용하고

2류 리더는 타인의 능력을 이용하고

1류 리더는 타인의 지혜를 이용한다.

• Some of cell death processes are programmed  programmed cell death
• Various cell death
• Apoptosis depends on an intracellular proteolytic cascade mediated by caspases
• Apoptosis depends on an intracellular proteolytic cascade mediated by caspases
• Extrinsic pathway: cell-surface death receptors
• Intrinsic (mitochondrial) pathway
• Bcl2 family proteins regulate intrinsic pathway of apoptosis
• Some Bcl2 family proteins are pro-apoptosis, while others are anti-apoptotic
• Extracellular survival factors inhibit apoptosis in various ways
• Apoptotic cells and their fragments remain intact and are phagocytosed, triggering no
inflammation

• Phosphatidylserine is normally located at the internal leaflet but exposed upon cell
death  ‘eat me’ signal

• Normal cells also have ‘don’t eat me signal’ such as CD47

• Decreased apoptosis contributes to tumor growth