Cell Cycle and Apoptosis
September 21, 2017
Control of the Cell Cycle
-The proteins that control
the cell cycle are different
from the proteins that are
involved in the process
-Series of checkpoints
-Each checkpoint serves as a
biochemical switch
Control of the Cell Cycle
The control system:
1. Swithes are binary:
“On or Off” and once started
the process continues to
completion
2. Reliable due to back-up
systems
3. Adaptable so it can be
modified to different cell
types or conditions
Key Control Components
1. Cyclin-dependent kinases (Cdk)
-Activities vary throughout the cell cycle
-The targets of phosphorylation then varies
2. Cyclins
-Control the activity of Cdk’s
-Undergo synthesis and degradation in
each cell cycle
Cycling of the cyclins creates the cycles of the cyclin/Cdk complex
allowing for the varying activities resulting in progression through
the cell cycle
Cyclin-Cdk Complexes
G1-cyclin
4 classes of cyclins
-All eucaryotes require 3 cyclins (G1/S, S, M)
Yeast – 1 Cdk
Vertebrates – 4 Cdks
Cyclin – activates the Cdk and helps direct it to the specific target
Cdk Activation
Cdk fullly active
1. Cyclin bound
2. Phosphorylated at the active site
Regulation of Cdk Activity
-Cdk activity can be fine tuned through 2 phosphorylations at
the top of the active site
-Important in control of M-Cdk activity
Inhibition of Cyclin-Cdk Complexes
-Binding of inhibitors alters the structure of the active site
-Cdk inhibitors are utilized more early in the cell cycle
Cyclin-Cdk Complexes
G1-cyclin
APC/C Anaphase-Promoting Complex, or Cyclosome
-Regulated by protein destruction
Marking Proteins by Ubiquitin
Ubiquitination Process I
Ubiquitination Process II
There are about 300 different E2-E3 each recognizing a different degradation
signal. Therefore subsets of proteins can be regulated as a group.
Control of Proteolysis of APC/C
-Degrading S and M-cyclins stops
the Cdk activity and the Cdk’s
or Cdh1 targets become dephosphorylated
and inactive
or S-cyclin -APC/C is active in G1 keeping
Cdks inactive
Control of Proteolysis by SCF
-Degrades Cdk Inhibitors of S-Cdks and allows S phase to occur
-F-box protein is constant through the cell cycle and is used to
recognize the target
Cell-Cycle Control Overview
10% of yeast genes encode mRNAs which oscillate in the cell cycle
Creation of DNA Lesions
“Endogenous”
Cellular Metabolism
Oxidation Hydrolysis Alkylation
8-hydroxyguanine FAPY Abasic site 3-methyladenine O6 -methylguanine
Radiation Chemotherapy
Therapy
“Exogenous”
DNA Damage Signaling Pathways
DNA Damage
Checkpoint
Mdm2 – ubiquitin ligase
p21 – CKI (Cdk Inhibitor Protein)
Check1/2
Chk1/2 phosphorylate Cdc25 thereby inactivating the
phosphatase activity resulting in inactive Cdk
DNA Damage
-Cell cycle is arrested until the damage is repaired
-If it’s not repaired:
1. Unicellular organisms will resume their cycle taking a
potential mutation over death
2. Multicellular organisms will sacrifice a cell over the
health of the organism
Apoptosis
Apoptosis
Apoptosis is one type of programmed
Process of Apoptosis
-Cells shrink and condense
-Cytoskeleton collapses
-Nuclear envelope disassembles
-Nuclear chromatin fragments
-Cell surface belbs and may break up (apoptotic bodies)
-Cell surface is chemically altered
-Macrophage engulfs the cell
Process of Cell Necrosis
-Cell insult or injury
-Cells swell and burst
-Cell contents are released
-Inflammatory response
Examples of Apoptosis
1. Number of cells
-Nervous system
Examples of Apoptosis
2. Development
Paws of embryonic mouse
Examples of Apoptosis
3. Regulate cell numbers
-Liver
4. Quality control
-Eliminates abnormal, misplaced, nonfunctional,
or dangerous cells
-Developing T and B cells that do not produce
useful antigen receptors or that are self-reactive
5. Supply of cells
-Large numbers of neutrophils are produced and
stored awaiting infection
Biochemical Characteristics
DNA Cleavage
-Endonuclease cleaves
DNA into fragments
between nucleosomes
Biochemical
Characteristics
TUNEL Assay - TdT-mediated
dUTP nick end labeling
Phosphatidylserine localization
-movement from the inner
to outer membrane
-marks the cell for
macrophages
Caspases
-Enzymes responsible for apoptosis
-A family of proteases that cleave proteins at aspartic acid
residues
-C for cysteine (in active site) and ASP for aspartic acid
-Synthesized as inactive precursors, procaspases
-Not all caspases are involved with apoptosis
-ICE, interleukin-1-converting enzyme
Procaspase Cleavage
Caspases
-Initiator procaspase, start the proteolytic cascade
-Executioner procaspases, cleave and activate other
executioner procaspases and other targets
-Targets include: nuclear lamins, endonuclease inhibitor,
cytoskeleton components, cell-cell adhesion proteins
-The caspase cascade is:
-Very destructive
-Self-amplyfing
-Irreversible
Caspase Cascade
Caspases
-Initiator caspases have a caspase recruitment domain (CARD)
that enables them to bind to adaptor proteins into activation
complexes
-In the complex, the initiator caspases are close enough to
activate each other starting the cascade
-2 Apoptotic pathways
1. Extrinsic Pathway
2. Intrinsic Pathway
Extrinsic Pathway
DISC – death-inducing signaling complex
Inhibitors such as decoy receptors and intracellular blocking proteins
Intrinsic Pathway
Releases mitochondrial proteins into
the cytoplasm
Cytochrome C release can trigger
apoptosis through interaction with the
adapter protein Apaf1
Intrinsic Pathway
Classes of Bcl2 Proteins
Bcl2 proteins –regulate apoptosis through controlling the release of cytochrome c
Pro-Apoptotic BH123
Regulation of
Intrinsic
Pathway
BH3-only proteins activate
apoptosis through direct
binding with anti-apoptotic
proteins
-p53 activates BH3-only
proteins (Puma and Noxa)
-Bid – extrinsic and intrinsic
pathways
Model for
IAPs
Inhibition of Apoptosis
(BH3-only
(Anti-apoptotic) pro-apoptotic)
