Lecture

Apoptosis

Dr. Shazia Rashid

Dr Shazia Rashid
 Apoptosis
• Programed death of cells to control excessive proliferation by eliminating
the “unnecessary”, mutated, or infected cells, the events are collectively
referred to as apoptosis
• it is a normal occurrence, neat and orderly process

• Greek word that means “dropping off” or “falling off,” as leaves from a
tree

• Apoptosis was first described by Kerr et al. (1972)

• Important for maintaining homeostasis in multicellular organisms and

failure to regulate apoptosis can result in serious damage to the organism,
causing diseases like cancer, Alzheimer’s and Parkinson’s

Dr Shazia Rashid
Dr Shazia Rashid
• Cell number is tightly regulated in multicellular organisms

• Controlled by rate of cell division and also by rate of cell death

• If cells are no longer needed, they commit suicide

Apoptosis- most common form of programmed cell death

Dr Shazia Rashid
Developing mouse paw
 Role of Apoptosis:
i) It has important role in various fundamental biological processes such
as organ development, wound healing, immune response and
oncogenesis.

ii) Plays important role in regulating total cell number.

iii) It helps in adaptation of an organism to environment and resolution of

inflammation by safe elimination of unwanted cells.

iv) It helps in removing the damaged, infected and potentially neoplastic

cells and thus protect the human beings and different livestock from
various diseases.

Dr Shazia Rashid
 Machinery responsible for apoptosis:

Caspase family of proteins:

• Cysteine proteases (proteases with a key cysteine residue in their catalytic

site)
• Caspases are present as inactive precursors, procaspases.
• Procaspases activated by proteolytic cleavage in response to signals that
induce apoptosis
• Most important player of apoptosis inducing most of the changes
Function:
• The activated caspases cleave, thereby activate other members of the
procaspase family resulting in an amplifying proteolytic cascade.
• They also cleave other key proteins in the cell e.g. One caspase cleaves the
lamin proteins resulting in the irreversible breakdown of the nuclear
membrane.

Dr Shazia Rashid
Dr Shazia Rashid
 Apoptosis can be triggered by:

1. Internal stimuli , such as abnormalities in the DNA, lack of oxygen,

extremely high concentration of Ca2+, viral infection, severe oxidative
stress – Intrinsic pathway

2. External stimuli, such as certain cytokines (proteins secreted by cells of the

immune system), ionising radiations, elevated temperature, viral infection,
toxic chemical agents – Extrinsic pathway

• The is a cross-talk between two pathways – extra-cellular apoptotic signal

can cause activation of the intrinsic pathway

Dr Shazia Rashid
 Extrinsic Pathway
• Originates in the cell exterior and is triggered by Tumor Necrosis Factor (TNF) –
ability to kill tumor cells.
• TNF produced by cells of immune system in response to adverse conditions, such
as exposure to ionising radiations, elevated temperature, viral infection, toxic
chemical agents
• Involves cell surface receptors (Fas, TNF-R,TRAIL-R), also called death receptors
that belong to TNF-Receptor family that turn on the apoptotic process
• Each TNF receptor has a cytoplasmic domain containing a segment of 70 amino
acids, called “death domain” that mediates protein-protein interactions
• Binding of TNF family members to the receptor on the cell surface trigger their
activation causing trimerization and conformational change of the death domain,
leading to recruitment of a number of proteins

Dr Shazia Rashid
• Activated death receptors recruits procaspase 8 which joins the complex
and induce its cleavage and is activated. Thus forming death inducing
signalling complex (DISC)

• Caspase-8 is described as an initiator caspase because it initiates

apoptosis by cleaving and activating downstream, or executioner
caspases, that carry out the controlled self-destruction of the cell

Dr Shazia Rashid
 And TRADD

Dr Shazia Rashid
 Intrinsic pathway
• Activated in response to cell-death signals originating from the cell interior
such as lack of oxygen, extremely high concentration of Ca2+, viral infection,
severe oxidative stress (production of large numbers of destructive free
radicals)
• Activation of the pathway regulated by members of Bcl-2 family of proteins
(Bcl-2 acts as an oncogene by promoting survival of potential cancer cells that
would otherwise die by apoptosis)
• Bcl-2 family divided into three groups:
1. Proapoptotic members that promote apoptosis (e.g. Bax ad Bak)
2. Anitiapoptotic members that protect cells from apoptosis (e.g. Bcl-xl, Bcl-w,
and Bcl-2)
3. BH3-only proteins promote apoptosis by indirect method (e.g. Bid, Bad,
Puma and Bim)

Dr Shazia Rashid
Apoptosis is promoted by either:
• By inhibiting antiapoptotic Bcl-2 members
• By activating proapoptotic Bcl-2 members

• In a healthy cell: BH3-only proteins are absent or strongly inhibited, and

the antiapoptotic Bcl-2 proteins are able to restrain proapoptotic
members. Balance of pro and anti.
• During stress: BH3-only proteins are expressed and activated, shifting the
balance towards apoptosis
• In these circumstances, the restraining effects of the antiapoptotic Bcl-2
proteins are overridden and Bax translocates from cytosol to the outer
mitochondrial membrane
• This increases the permeability of the outer mitochondrial membrane and
promotes the release of cytochrome c (resides in intermembrane space),
this is the “point of no return”-irreversible commitment of cell to
apoptosis

Dr Shazia Rashid
• In the cytosol, cytochrome c forms a multiprotein complex with the
cytoplasmic protein, Apoptotic protease activating factor 1(Apaf-1) and
procaspase-9 form the apoptosome

• This complex activates caspase-9 (initiator caspase), which in turn

activates downstream executioner caspase which bring about apoptosis

Formation
Dr Shazia Rashid of Apoptosome
Dr Shazia Rashid
• The activation of the caspase cascade can be blocked by inhibitors of
apoptosis proteins (IAPs), which can be upregulated in response to
survival signals.

Dr Shazia Rashid
Cross talk between the Intrinsic and
Extrinsic pathways

Dr Shazia Rashid
Cross talk between the extrinsic and
intrinsic pathways
– Cross talk between the extrinsic and intrinsic pathways is mediated by
Bid, a proapoptotic Bcl-2 family member.

– Caspase-8 mediated cleavage of Bid which increases its activity and

results in translocation to the mitochondria, where it acts together
with two other proapoptotic proteins, Bax and Bak, to induce
cytochrome c release, thereby activating the intrinsic pathway.

Dr Shazia Rashid
 Cell Necrosis
• Cell death due to response to tissue injury
• Cells swell by absorbing water and burst, releasing their intracellular
contents, which can damage surrounding cells and frequently cause
inflammation.

Causes of necrosis:
(i) Micro-organism and their Products: Invasion of cells by different micro-
organisms, such as Mycobacterium tuberculosis, Corynebacterium
diphtheriae and toxins of Clostridium sp.
(ii) Physical Agents: Various physical agents like electricity, extreme heat and
cold, X-rays and prolonged pressure (by ligature or tumours).
(iii) Chemical Agents: Various chemicals such as carbolic acids, mineral acids
and caustics act directly on cells resulting into necrosis of cells.

Dr Shazia Rashid
Difference between Apoptosis and
necrosis?

Dr Shazia Rashid
• Differentiate between apoptosis and necrosis?

Dr Shazia Rashid