Role of NF-B in the

Regulation of Immunity
and Apoptosis

Kavitha Bharatham
 Abstract

Introduction
Structural Description
Role as regulator of Immune responses
Role as regulator of apoptosis
Regulation Mechanism
As target for Therapy
Conclusions
Introduction
 Nuclear Factor kappa B (NF- B)
Multifunctional transcription factor

Play central role in the cellular response to a variety of stress

signal by regulating genes

Exists as homo or hetero-dimers found inactive in the

cytoplasm

Upon stimulation, active NF-B rapidly translocates to the

nucleus where it binds B-sites and activates target genes
 Role of NF- B/Rel family
Involved in proinflammatory response: a first line of defense
against infectious diseases and cellular stress
Signal Activated NF- B immune defence activated
Immune response, inflammatory response, accute phase
response
NF-B also a major anti-apoptopic factor
aberrant activation of NF- B causes
Inflammatory diseases, Rheumatoid arthritis,
Asthma, Atherosclerosis, Alzheimer
helps keeping cancer cells alive
NF-B also promoting growth
Activated NF- B cyclin D expression enhanced growth
Factors that induce NF-kB

Reactive Oxygen
Species (ROS)

Leukemia 16:1053-1068, 2002

 Genes Regulated by NF-kB

Nat.Rev.Cancer 2:301-310, 2002

 Disorders associated with aberrant
NF-B activation
Rheumatoid arthritis Autoimmune thyroid
Atherosclerosis disease
Vascular dysfunction Cystic fibrosis
Multiple sclerosis Diabetes
Neurodegenerative disorders Aging
Inflammatory bowel disease Macular degeneration
H. pylori-associated gastritis HIV/AIDS
Systemic inflammatory Cancer
response syndrome Septic shock

And the list is growing

Structural Description
 The NF- B/Rel family
NF-B belongs to the Rel family, which contains five
mammalian Rel/NF- B proteins:
Potent transactivators synthesized in their mature form
1. RelA (p65)
2. c-Rel
3. RelB
Synthesized as precursors that are post translationally processed with no
transactivation properties
1. NF- B1 (p105 undergoes proteolytic maturation to p50)
2. NF- B2 (p100 undergoes proteolytic maturation to p52)

RelB forms dimer only with p50/p52

Most Common form: p50/p65 (NF-B1/RelA)
 NF- B proteins structure

NH2 Rel homology domain GGG C-terminal IkB-like domains

p105
- TAD

p50 Ankyrin Repeats

p100

p52

RelA(p65)
+TAD

cRel TransActivation
domains (TAD)
RelB
 Structural Details
Rel Homology Domain (RHD): 300aa conserved domain of
Rel family which contains NLS (Nuclear localisation sequence)
with several functions
DNA-binding (N-terminal half)
Dimerization (C-terminal half)
IB-interaction (C-terminal half)

IB (Inhibitor of Nf- B) blocks NLS and abolish

translocation to nucleus
IB family includes IB-, IB-, IB- and IB-, Bcl-3, etc.,
They posess ankyrin-repeats which are necessary for RHD-interaction
impedes DNA-binding
Upon Signal undergoes dissociation and degradation
 Dimerization

Dimer formation is necessary

for DNA-binding
Each subunit interacts with one
half site
B-sites are symmetric:
5-GGGRNNYYCC-3

p50
Structure: NFkB (p50-p65) + DNA

Side view

b
--5-GGGRNNYYCC-3--
- 3-CCCYNNRRGG-5--
Role as Regulator of
Immune responses
 Role in inflammation
TNF-alpha
Inflammation is a
process by which
the body attempts to
dilute, destroy, or
isolate a noxious
(harmful) agent and
repair damage

Transcription
TNF
Il-1
Up regulation of adhesion molecules (ICAM-1, VCAM-1)
Cytokines that further enhance the immune response
activators of inflammatory pathways (arachidonic acid met
abolites, superoxides and nitric oxide)
 Role of IKK in Activating NF-B
Pro-inflamatory cytokines,
Pathogen associated molecular patterns (PAMPs), TNF Receptor (TNFR),
Toll-like Receptor (TLR), Interleukin-1 receptor (IL-1R)

Two types of inactive

complexes in the cytoplasm
Trimers = RHD-Homo-or
heterodimers bound to an IB-
repressor
Heterodimers = Rel-protein + Transcription
unprocessed RHD-precursor
(p105, p110)
NLS is exposed and B site
translocated to
Nucleus
 Two main signaling pathways
Innate Adaptive

NUCLEUS
mRNA
Target genes Transcription
 Innate vs Adaptive
NF-kB activation pathway
Canonical/Classical/Innate NonCanonical/NonClassical/Adaptive
NF-B activation pathway NF-B activation pathway

Applies to RelA-p50 and c-Rel-p50 Affects NF-B2, which preferentially

Retained in cytoplasm by IB dimerizes with RELB

Triggered by microbial and viral Triggered by members of the tumor-

infections and exposure to proinfl necrosis factor (TNF) cytokine family
ammatory cytokines

Depends mainly on the IKK sub Depends selectively on activation of the

unit of the IKK complex. IKK subunit + another kinase NIK.

Induce the phosphorylation-depen Induce the phosphorylation-dependent

dent proteolytic removal of the I proteolytic removal of the IB-like
B C-terminal domain of NF-B2
Role as Regulator of
Apoptosis
 Apoptosis is Programmed Cell Death
Apoptosis is needed to destroy cells
that represent a threat to the integrity
of the organism
Inducers:
Damage-related inducers
heat shock, viral infection, bacterial toxins,
oncogenes (myc, rel, E1A), tumor
suppressors (p53), cytolytic T cells,
oxidants
Therapy-associated agents
Chemotherapeutic drugs (e.g., cispatin,
nitrogen mustard)
Antracyclines (doxorubicin), gamma
radiation, UV radiation
Toxins
Ethanol, -amyloid peptide
 Diseases Associated with Deregulated
Apoptosis
Increased Apoptosis
AIDS
Neurodegernative disorders
Alzeheimers disease,
Parkinsons disease,
Amyotrophic lateral sclerosis
Retinitis pigmentosa
Myelodysplastic syndromes
Aplastic anaemia
Ischaemic Injury
Myocardial infarction,
Stroke,
Reperfusion injury
Toxin-Induced liver disease
Alcohol
Inhibition of Apoptosis
Cancer
Follicular lymphomas
carinomas with p53 mutations
hormone dependent tumours:
breast cancer, prostate cancer,
ovarian cancer
Autoimmune Disorders
Systemic lupus erythematosus
Immune-mediated glomerulonephritus
Viral Infections
Herpesvirus, poxvirus, adenovirus
NF-B is an anti-apoptotic factor

Via NF-B
TNF blocks its own cell death potential

Chemotherapy activates NF-B

within tumor cells

NF-B inhibitors
augment chemotherapy
Mechanism behind Anti-apoptotic
activity
RIP: Receptor interacting protein
FADD: Fas associated Death domain
IAP: Inhibitor of Apoptosis
TRAF: TNFR associated protein
Bid: Bcl-2 interacting domain
JNK: Jun N-terminal kinase

Cell Death
Balance between life and death
When NF-B is not When NF-B is
inhibited inhibited

ANTI-APOPTOTIC PRO-APOPTOTIC
PROTEINS PROTEINS
Regulation Mechanism
 Negative feedback: Attenuation of
NF-B response
Negative loop: IB is under direct control of NF-B
IB

IB
RelA
ib p50
kB site

RIP and TRAF1 are cleaved

Caspase 8
RIP Cleaved
TRAF1
Therapeutic inhibition of NFB
 Conclusions
The NF-B is an important pathway in regulating the stress
response in the body
It plays a key role in oncogenesis
Work continues on manipulating the pathway for use in
therapy
Complete elucidation of the mechanisms involved in
regulation of NF-B activation is required to generate
inhibitors
Therapeutic inhibitors that selectively block NF-B activation
in cancer cells without effecting normal functions are required
 References
Seminars in Cancer Biology 13 (2003) 107114
TRENDS in Immunology Vol.25 No.6 June 2004
Clinical Chemistry 45:1 717 (1999)
The Journal of Clinical Investigation January 2001 Volume 107 Number 2
Human Molecular Genetics, 2002, Vol. 11, No. 20