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Cellular and Pharmacological
signaling through receptors

Intracellular Receptors:

‡ Nuclear receptors: ligand-activated

transcription factors that activate mRNA
synthesis

Membrane receptors:

Three main types of membrane receptors

‡Tyrosine Kinases
‡G-protein linked
‡Ion channels
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Nuclear Receptor structure-function«..
Receptor protein homologies reflect function &
diversity of ligands

steroids
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˜teroid receptors:
Glucocorticoid
Mineralocorticoid
Progesterone
Androgen
Estrogen

Others *:
T3R
RAR
VD3R
PPARs

* The RXR is a common partner for those in

the second group
Öow do nuclear receptors activate
specific genes??

They recognize specific sequences of

DNA called ³hormone response elements´
which are a class of ³enhancers´

Usually there are two identical or similar

parts to the ÖRE called ³half
³half--site motifs´
that each bind to a nuclear receptor.
Therefore nuclear receptors bind as dimers.
promoter
gene target

enhancer
]hat structural features of nuclear receptors
interact with (bind to) DNA??
Ȣ 

È
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Nuclear receptor ³Zn fingers´

P = proximal
D = distal
 

 


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ë) P box ï ëo nucleotide sequence

2) D box ï spacing of the half-site motif

On the basis of the primary nucleotide recognition

sequence nuclear receptors can be divided into
two families:

ë) glucocorticoid-R family
2) estrogen/thyroid Öormone-R family
Öomology between the P and D boxes
of nuclear receptors


amino acid sequence is the same for the ER, TR

RAR, AR etc

È
amino acid sequence is different in all these nuclear

receptors
˜ince the nuclear receptor Zn2+ finger
structures are all different what do the DNA
response elements for each nuclear receptor
look like ?
] 

   
 $
˜urvey of the elements required for gene
activation
ï promoter analysis (ÖRE)
(enhancer)

consensus sequence.

Glucocorticoid receptor (GRE)

G GTACAN N NTGTTC T

Estrogen receptor (ERE)

T
GGTCAN NNTGAC C
palindromic
Consensus sequences for enhancers which
bind and are transactivated by the ER/TR
family:

Öalf-site motif: A G G T C A as a repeated

sequence

N pN pN p Vit D3

NpNpNpNp T3

NpNpNpNpNp RAR
Öow do nuclear receptors activate transcription?
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Co-repressor complex keeps the gene silent when there

Is no ligand around

Target gene
 ]hat does Öistone Acetylation accomplish?

ë) Acetylation neutralizes the positive charge on histone tail to

³loosen´ the interaction between DNA and the histone

2) Acetylation patterns control the binding of nonhistone proteins

(ie. like the RNA polymerase complex) to the chromatin fiber.
 ÖAT

(
!&&
)( RNA
Polymerase
pë

'( complex
OAc-
RAR RXR
Target gene
AGGTCANNNNN AGGTCA promoter
DNA response element
histones

ë) The binding of ligand (RA) induces a conformational

change in the RAR/RXR heterodimer
2) Recruitment of pë
and p3

ÖAT proteins
histone acetylation opens the DNA.
3) This open conformation allows the RNA polymerase complex
proteins access to the promoter to allow transcription
of mRNA to occur.
There are natural and synthetic
ligands for nuclear receptors
 ˜ynthetic ligands

˜elective Estrogen Receptor Modulators (˜ERM˜)

˜ERMs are used in the treatment of breast cancer

 ˜elective Estrogen Receptor Modulators
(˜ERM˜)

ï A need for safe hormone replacement

therapy (ÖRT)
ï breast cancer prevention & treatment

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The ³biocharacter´ of ˜ERMs is dependent on
several factors
˜ERMs will elicit different effects depending on
several factors in different tissues and on different
gene enhancers (response elements)

˜ERMs differentially affect the interaction of

the ER with coregulators
Relative levels of coactivators in a tissue type
may determine agonist/antagonist

 ˜ERMs will differentially activate gene

transcription depending on the DNA sequence
of the response element.
 ˜ERM biocharacter depends on the subtype of ER

There are two ER subtypes: ERĮ and ERȕ

Tam ï bound to ERĮ is sometimes an agonist

sometimes an antagonist
Tam ï bound to ER
is always an antagonist
]hen both ER subtypes are expressed in the same cell
the combined response is different from individual
responses.
  

 ! 

Retinoic acid & derivatives are:

cancer (a) anti-proliferative (prevent growth)

therapy (b) - differentiation inducing (very important
during
development)
- morphogenic

³Promyelocytic´ leukemia
ï RA induces terminal differentiation into
myelocytes
  ! 

Retinoids can reduce wrinkles and reduce acne:

ë) inhibit metalloproteinases that breakdown collagen
2) induce cell proliferation under some circumstances
3) cause epidermal differentiation and drying
 Peroxisome Proliferators

PPs comprise a diverse group of non-genotoxic carcinogens

including industrial plasticizers, hypolipidemic drugs, herbicides
and some leukotriene antagonists. As their name implies,
these chemicals are known to cause a marked increase in
size and abundance of hepatocyte peroxisomes when administer
-ed to rats and mice.
Peroxisomes are single membrane cytoplasmic organelles,
designed to metabolize long chain fatty acids through oxidase
enzymes. In coincidence with proliferation of these organelles,
PPs induce many peroxisomal and mitochondrial enzymes
involved in fatty acid ȕ-oxidation, cytochrome P45
fatty acid
oxidation and cholesterol homeostasis.

 


* 
 
 #!      !  

-fatty acid sensors

-act to control networks of target genes involved in
energy metabolism

ë) PPAR-Ä is the master regulator in the formation of fat

cells and their ability to function normally in the adult
-Lipids in the circulation result in insulin resistance therefore
this receptor is an important regulator of glucose metabolism
-insulin-sensitizing TZD drugs bind to PPAR-Ä
-found predominantly in the liver

2) PPAR- is activated during prolonged fasting (such as

during the night) when fatty acids are released from adipose
and transported to the liver for oxidation and production of
energy
-induction results in improved ÖDL profiles (³good cholesterol´)
-ligands include the group of fibrate drugs used to treat
hyperlipidemia
 Cell. 2

3 Apr Related Articles, Links

ëëë3(2):ë59-7
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.,

.

Gene Expression Laboratory, The ˜alk Institute, ë

ë

North Torrey Pines Road, La Jolla, CA 92
37, U˜A.
3) PPAR-Ö is ubiquitously expressed and is a strong
regulator of fatty acid oxidation and energy homeostasis.
-˜ynthetic ligand G]5
ë5 lowers plasma triglycerides
and increases ÖDL levels
-transgenic mouse expressing an activated form of PPAR-Ö
in adipose tissue is lean and is resistant to obesity and
hyperlipidemia
-PPAR-į is a major fat burning regulator that opposes the
activity of PPAR-Ä
-Transgenic mouse expressing PPAR-Ö in skeletal muscle
dramatically increases non-fatiguing type I fibers to produce
a mouse capable of running much longer than the control
mouse without rest (³marathon´ or ³Lance Armstrong
Mouse´)
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Note:
PPAR ±gamma Ȗ is slightly increased in obese patients
Receptor Ligands Target genes
(natural and synthetic)
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