APOPTOSIS: An overview
Sanjeev Sharma*, Aarti Bhardwaj$, Shalini Jain# and Hariom Yadav#
*Animal Genetics and Breeding Division, #Animal Biochemistry Division, National Dairy Research Institute, Karnal-132001, Haryana, India $College of Applied Education and Health Sciences, Meerut, U.P.
INTRODUCTION
Cell death by injury
-Mechanical damage -Exposure to toxic chemicals
Cell death by suicide
-Internal signals -External signals
Conted..
Apoptosis or programmed cell death, is carefully coordinated collapse of cell, protein degradation , DNA fragmentation followed by rapid engulfment of corpses by neighbouring cells. (Tommi, 2002)
Essential part of life for every multicellular organism from worms to humans. (Faddy et al.,1992) Apoptosis plays a major role from embryonic development to senescence.
Why should a cell commit suicide?
Apoptosis is needed for proper development
Examples:
The resorption of the tadpole tail
The formation of the fingers and toes of the fetus The sloughing off of the inner lining of the uterus The formation of the proper connections between neurons in the brain
Apoptosis is needed to destroy cells
Examples:
Cells infected with viruses Cells of the immune system Cells with DNA damage
Cancer cells
What makes a cell decide to commit suicide?
Withdrawal of positive signals
examples : growth factors for neurons Interleukin-2 (IL-2)
Receipt of negative signals
examples : increased levels of oxidants within the cell damage to DNA by oxidants death activators : Tumor necrosis factor alpha (TNF-) Lymphotoxin (TNF-) Fas ligand (FasL)
History of cell death / apoptosis research
1800s 1908 Numerous observation of cell death Mechnikov wins Nobel prize (phagocytosis)
1930-40
1948-49 1955
Studies of metamorphosis
Cell death in chick limb & exploration of NGF Beginning of studies of lysomes
1964-66
1971 1977
Necrosis & PCD described
Term apoptosis coined Cell death genes in C. elegans
1980-82
1989-91
DNA ladder observed & ced-3 identified
Apoptosis genes identified, including bcl-2, fas/apo1 & p53, ced-3 sequenced
(Richerd et.al., 2001)
NECROSIS Vs APOPTOSIS
Wilde, 1999
Necrosis vs. Apoptosis
Necrosis
Cellular swelling Membranes are broken ATP is depleted Cell lyses, eliciting an inflammatory reaction DNA fragmentation is random, or smeared In vivo, whole areas of the tissue are affected
Apoptosis
Cellular condensation Membranes remain intact Requires ATP Cell is phagocytosed, no tissue reaction Ladder-like DNA fragmentation In vivo, individual cells appear affected
STAGES OF APOPTOSIS
Induction of apoptosis related genes, signal transduction
Sherman et al., 1997
APOPTOSIS: Morphology
organelle membrane blebbing & changes mitochondrial leakage
reduction
cell
shrinkage
nuclear fragmentation
chromatin condensation
Hacker., 2000
APOPTOSIS: Morphological events cell shrinkage
organelle reduction mitochondrial leakage
chromatin condensation nuclear fragmentation membrane blebbing & changes
Blebbing & Apoptotic bodies
Bleb The control retained over the cell membrane & cytoskeleton allows intact pieces of the cell to separate for recognition & phagocytosis by MFs
Apoptotic body
MF
MF
Caenorhabditis elegans
1090 cells
decision to die
131 cells
execution
engulfment
apoptosis
degradation
ced-3 ced-4
egl-1
ced-9
ced-1 ced-2 ced-5 ced-6 ced-7 ced-10
nuc-1
ces-2
ces-1
Apoptosis: Pathways
Extrinsic Pathway
Death Ligands Death Receptors
Initiator Caspase 8
Effector Caspase 3
Intrinsic Pathway
PCD
DNA damage & p53
Mitochondria/ Cytochrome C
Initiator Caspase 9
MAJOR PLAYERS IN APOPTOSIS
Caspases Adaptor proteins TNF & TNFR family Bcl-2 family
Ligand-induced cell death
Ligand FasL TNF TRAIL Receptor Fas (CD95) TNF-R DR4 (Trail-R)
Ligand-induced cell death
The death receptors
FasL Trail
Ligand-induced trimerization
TNF
Death Domains Death Effectors Induced proximity of Caspase 8
Activation of Caspase 8
APOPTOSIS: Signaling & Control pathways I
Externally driven Externally driven
Apoptotic signals
p53 Internally Cytochrome C driven
mitochondrion
Activators of initiator enzymes
Initiator caspases
6, 8, 9,12
Execution caspases
2, 3, 7
Apoptosis events
Activation
APOPTOSIS: Signaling & Control pathways II
Externally driven Externally driven
Apoptotic signals
p53 Internally Cytochrome C Bcl2 driven
Inhibitors Activators of initiator enzymes
External Internal
Initiator caspases
6, 8, 9,12
Survival factors
Execution caspases
2, 3, 7
Apoptosis events
Inhibitors of apoptosis
Inhibition
The mitochondrial pathway
DNA Fas damage Casp8 p53 Bid Bid Bid Bax
Growth factor receptors
PI3K
BAD Bcl2
Akt
casp3
Bax
casp9 Apaf1
ATP Cyt.C Smac/ DIABLO
IAPs casp3
H2O2
AIF
Pollack etal., 2001
REGULATION OF APOPTOSIS
Stimuli apoptosis selection of targets
(Rich et al., 2000)
Apoptosis by conflicting signals that scramble the normal status of cell (Canlon & Raff, 1999) Apoptotic stimulicytokines, death factors (FasL)
(Tabibzadeh et al., 1999)
DNA breaks p53 is activated arrest cell cycle or activate self destruction (Blaint & Vousden, 2001)
Importance of Apoptosis
Important in normal physiology / development
Development: Immune systems maturation, Morphogenesis, Neural development Adult: Immune privilege, DNA Damage and wound repair.
Excess apoptosis
Neurodegenerative diseases
Deficient apoptosis
Cancer Autoimmunity
FUTURE PERSPECTIVES
The biological roles of newly identified death receptors and ligands need to be studied
Need to know whether defects in these
ligands and receptors contribute to disease
CONCLUSION
an important process of cell death can be initiated extrinsically through death ligands (e.g. TRAIL, FasL) activating initiator caspase 8 through induced proximity. can be initiated intrinsically through DNA damage (via cytochrome c) activating initiator caspase 9 through oligomerization. Initiator caspases 8 and 9 cleave and activate effector caspase 3, which leads to cell death.
DNA DAMAGE
p53
The bcl-2 family
N
BH4
Raf-1 calcineurin
BH3
BH1
BH2
TM
Receptor domain Ligand Pore domain formation phosphorylation
Membrane anchor
Group I Group II
Bcl-2 bax Bad bid bik
Back
Group III
P53 & Apoptosis
p53 first arrests cell growth between G1 S This allows for DNA repair during delay
If the damage is too extensive then p53 induces gene activation leading to apoptosis (programmed cell death)
BACK
3 mechanisms of caspase activation
a. Proteolytic cleavage e.g. pro-caspase 3
b. Induced proximity, e.g. pro-caspase 8
c. Oligomerization, e.g. cyt c, Apaf-1 & caspase 9
Back
Apoptosis signal to kill infected cells
Cytolytic lymphocyte/CTL (& natural killer lymphocyte) presents Fas ligand/CD178 on its surface to tell the infected cell to die
Fas ligand
Externally driven
Apoptotic signals
Cytochrome c
Initiator caspases
The immunological synapse holds the cells much tighter together than shown here Fas/ CD95 is the death receptor
Execution caspases
Apoptosis events
