Advanced Cell Biology

Department of Biology and Biotechnology

Faculty of Science
The Hashemite University

Prof. Lubna Tahtamouni

Spring, 2022/2023
 Module 5
Cellular Communication
• Cell signaling: the basics
• Cell surface receptors: G-protein coupled receptors, Enzyme coupled
receptors
• Gene expression regulation by proteolysis
All cells communicate!
 Budding yeast Saccharomyces cerevisiae: a well-
studied model system
• Unicellular eukaryotes communicate (e.g. yeast mating).
• Animals have more complex signaling systems.

Figure 15-2 Molecular Biology of the Cell (© Garland Science 2008)

Higher eukaryotic cells have complex signaling pathways
General model for a signaling pathway
• Communication between cells – extracellular signaling
molecules.

 Membrane receptors
General model for a signaling pathway
• Communication between cells – extracellular signaling
molecules.
 Extracellular signaling molecules bind to specific
receptors
• Different kinds of signaling molecules: proteins, nucleotides,
steroids, etc.
• Signal-receptor complexes initiate a response.

Figure 15-3 Molecular Biology of the Cell (© Garland Science 2015)

 Receptors are “Signal transducers”
Convert an extracellular ligand binding event into
intracellular response
Extracellular signal molecules can act over short (a,b,c)
or long distances (d)
Autocrine signaling
 Response to extracellular signals (change in
cell behavior) can be slow or rapid
* Changes in proteins already present can be fast.
 Delivery of signals could be slow or fast
• Endocrine signaling is relatively slow since signal molecules (hormones) are diluted
while being transported or require gene expression.
• Synaptic signaling is fast (milliseconds). Molecules in local high concentration.

Figure 15-5 Molecular Biology of the Cell (© Garland Science 2008)

 Cells are programmed to respond to
combinations of extracellular signals
• Each cell type displays a particular set of receptors.
 Different types of cells may respond differently to
the same extracellular signal molecule
• The type of response is determined by the machinery that interprets the
signal (Intracellular signaling proteins and effectors).
 Extracellular signaling molecules bind to specific
receptors

Figure 15-3 Molecular Biology of the Cell (© Garland Science 2015)

 Some signal molecules can diffuse into the cell
and activate intracellular receptors.
Example I: the gas Nitric Oxide (NO)
Some signal molecules can diffuse into the cell
and activate intracellular receptors.
Example II: ligands of nuclear receptors: Modulate transcription
of specific genes
Some signal molecules can diffuse into the cell
and activate intracellular receptors.
Example II: ligands of nuclear receptors: Modulate transcription
of specific genes
Three classes of cell
surface receptors
 Many intracellular signaling proteins function as
molecular switches that are activated by phosphorylation
or GTP binding
Many intracellular signaling proteins function as molecular
switches that are activated by phosphorylation
Many intracellular signaling proteins function as molecular
switches that are activated by GTP binding (G proteins)
Feedback loops in intracellular signaling
 Cell Signaling - Summary
• Each cell responds to a limited set of signals;
this depends on the set of receptors it has!
• These signals change the activity of internal
cellular proteins which changes the behavior of
the cell.
• These signals follow a chain of events known as
the signal transduction or pathway:
– A system of relaying information from the site
of reception to the point of action
– Normally the signal is amplified too - a small
input is quickly converted to a large response.
 Module 5
Cellular Communication
• Cell signaling: the basics
• Cell surface receptors: G-protein coupled receptors, Enzyme coupled
receptors
• Gene expression regulation by proteolysis
Three classes of cell surface
receptors
 G-protein-coupled receptors (GPCR)
• Largest family of cell surface receptors
• More than 800 members in human
• Multipass transmembrane proteins that has 7 α helices: N-terminal binds the ligand, C-
terminal binds G-protein
• Ligands: aa, small peptides, FA, odorants, photons….
• Receptors for smell, taste, sight and most cellular
responses
• Use G proteins to relay signal
• Activates enzymes, ion channels
• Target of half of all known drugs

Figure 15-21 Molecular Biology of the Cell (© Garland Science 2015)

 G-protein-coupled receptor (GPCR)
• Largest family of cell surface receptors
• More than 800 members
• 7-pass transmembrane proteins
• Use G proteins (trimeric proteins) to relay signal
• Target of half of all known drugs
 Structure of an inactive G protein
• They are trimeric (α,β,γ)
• Inactive when GDP is bound, active when GTP is bound
• Both alpha and gamma subunits are peripheral proteins attached to the membrane
through a lipid group.
• Beta and gamma subunits act as a unit.
 Structure of an inactive G protein
• They are trimeric (α,β,γ)
• Inactive when GDP is bound, active when GTP is bound
• The α subunit is a molecular switch GTPase. It switches between the
inactive GDP-bound form and the active GTP-active form.
Activation of a G
protein by an activated
GPCR
• Ligand binding to GPCR
activates it by inducing a
conformational change
• GPCR activation induces the α
subunit to release GDP and to
bind GTP
• Active GPCR acts as guanine
exchange factor (GEF)
• Active G proteins (α subunit,
and the β subunits as one unit)
regulate enzymes or ion
channels.
 Activation of a G protein by an activated GPCR
• Ligand binding to GPCR activates it by inducing a conformational change
• GPCR activation induces the α subunit to release GDP and to bind GTP
• Active GPCR acts as guanine exchange factor (GEF)
• Active G proteins (α subunit, and the β subunits as one unit) regulate enzymes or ion
channels.
• Once the job is done, GTPase function of the α subunit is stimulated by a specific
regulator of G-protein signaling (RGS or GAP).
 Effectors of G proteins
• Gαs: activates Adenylyl cyclase
• Gαi: inhibits Adenylyl cyclase
• Gαq: activates phospholipase C
• Gα12/13: activates GEF for Rho family of GTPases
 Effectors of G proteins
• Gαs: activates Adenylyl cyclase
• Gαi: inhibits Adenylyl cyclase
• Gαq: activates phospholipase C
• Gα12/13: activates GEF for Rho family of GTPases
Some G proteins (Gs) regulate the production of
Cyclic-AMP
• Synthesized by adenylyl cyclase and degraded by a
phosphodiesterase.
Some G proteins (Gs) regulate the production of
Cyclic-AMP
• Synthesized by adenylyl cyclase and degraded by a
phosphodiesterase.
 Cyclic-AMP-dependent protein kinase (PKA)
mediates most effects of cyclic-AMP
• It phosphorylates different target proteins according to the cell type.

(releases the regulatory units from catalytic units)

 Effectors of G proteins
• Gαs: activates Adenylyl cyclase
• Gαi: inhibits Adenylyl cyclase
• Gαq: activates phospholipase C-β
• Gα12/13: activates GEF for Rho family of GTPases
 Some G (Gq) proteins regulate the plasma
membrane-bound enzyme phospholipase C-β

q
 Some G proteins (Gq) regulate the plasma
membrane-bound enzyme phospholipase C-β
• It generates IP3 (soluble) and diacylglycerol (membrane bound)
Some G proteins regulate the plasma membrane-
bound enzyme phospholipase C-β
• Ca2+ release from the ER and activation of PKC at the plasma
membrane
 Ca2+ functions as an intracellular mediator
• Ca2+ acts as a signal: [Ca2+] ↓ in the cytosol but ↑ outside the cell
and in the ER.

Figure 15-41a Molecular Biology of the Cell (© Garland Science 2008)

 Calmodulin helps to relay the cytosolic Ca2+
signal
• CaM-kinase is switched on by Ca2+/calmodulin
• Then CaM-kinase phosphorylates itself (fully active) and other
proteins
• Ca2+ release from the ER activates NO synthase
 Effectors of G proteins
• Gαs: activates Adenylyl cyclase
• Gαi: inhibits Adenylyl cyclase
• Gαq: activates phospholipase C
• Gα12/13: activates GEF for Rho family of GTPases
Some G proteins (G12/13) activate Rho
GTPases GEF
Some G proteins (G12/13) activate Rho
GTPases GEF
Smell and vision depend on GPCRs that regulate
cyclic-nucleotide-gated ion channels
• Olfactory GPCRs → bind odors → [cAMP] ↑ → opens cAMP-gated
sodium channels → nerve impulse.
Smell and vision depend on GPCRs that regulate
cyclic-nucleotide-gated ion channels
• Activation of Rhodopsin (GPCR) is induced by light → activates cGMP
phosphodiesterase → [cGMP] ↓ → cation channels close →
neurotransmitter release ↓
Amplification in the GPCR pathways
Amplification in the GPCR pathways
 Target cells use 5 ways to become desensitized to an
extracellular signal molecule
1) Receptor sequestration
2) Receptor downregulation
3) Receptor inactivation
4) Inactivation of a signaling molecule
5) Introduction of inhibitory protein
 Regulation of GPCR (desensitization)
1) Receptor internalization
- Degradation by the lysosome
- Sequestration
2) Receptor inactivation (inhibiting GPCR from binding to G proteins)
• Receptor desensitization depends on receptor phosphorylation and arrestins
• Arrestin prevents interaction with G proteins.
• Arrestin mediates GPCRs endocytosis.

Or PKA or PKC (feedback

inhibition)
 Regulation of GPCR (desensitization)
1) Receptor internalization
- Degradation by the lysosome
- Sequestration
2) Receptor inactivation (inhibiting GPCR from binding to G proteins)
• Receptor desensitization depends on receptor phosphorylation and arrestins
• Arrestin prevents interaction with G proteins.
• Arrestin mediates GPCRs endocytosis.
 Module 5
Cellular Communication
• Cell signaling: the basics
• Cell surface receptors: G-protein coupled receptors, Enzyme coupled
receptors
• Gene expression regulation by proteolysis
Some subfamilies of receptor tyrosine kinases (RTKs)
 Some subfamilies of receptor tyrosine kinases (RTKs)
• RTKs are the most numerous enzyme-coupled receptors (~ 60
genes).
 Activation of receptor tyrosine kinases (RTKs)
• Ligand binding → receptor dimerization.
• Trans-autophosphorylation on tyrosine residues → higher activity.
• Docking sites for intracellular signaling proteins that has the SH2
domain or PTB domain
 Activation of receptor tyrosine kinases (RTKs) leads to
docking of SH2- or PTB-containing proteins
• Docking sites for intracellular signaling proteins that has the SH2
domain or PTB domain
• Examples:
Adaptor proteins, PLC, PI3K, Src
 Activation of receptor tyrosine kinases (RTKs) leads to docking
of SH2- or PTB-containing proteins
• Docking sites for intracellular signaling proteins that has the SH2
domain or PTB domain
• Examples:
Adaptor proteins, PLC, PI3K, Src
 Ras is a monomeric GTPase that relays signals
from RTKs

1) Ligand binds RTK

2) RTK dimerization, autophosphorylation
and activation
3) Docking of adaptor protein (Grb2)
containing SH2 domain
4) Binding of Ras GEF to Grb2, activation
of GEF
5) GEF stimulates the exchange of GDP
to GTP on Ras
6) Ras is active and activates effectors
Ras is a monomeric GTPase that relays signals from RTKs
• Molecular switch.
• Adaptors and GEFs couple RTK activation to Ras activation.
• Cell proliferation and differentiation.
• 30% of tumors have Ras mutations (rat sarcoma).
 Ras activates a mitogen-activated protein (MAP) kinase
module
• Activates transcription of specific genes such as cyclins (cell division).
Rho GTPases (Rho, Cdc42, Rac) relay signals from
RTKs
• Molecular switch.
• Adaptors and GEFs couple RTK activation to Rho/Rac/Cdc42 activation.
• Actin and microtubule cytoskeleton
The Rho family of GTPases couple cell-surface receptors
to the actin and microtubule cytoskeletons
 Activation of receptor tyrosine kinases (RTKs) leads to
docking of SH2- or PTB-containing proteins
• Docking sites for intracellular signaling proteins that has the SH2
domain or PTB domain
• Examples:
Adaptor proteins, PLC, PI3K, Src
 Phosphoinositide 3-kinase (PI 3-kinase) produces lipid
docking sites in the plasma membrane
• PIP3 remains in the membrane and is bound by various signaling proteins.
Phosphoinositide 3-kinase (PI 3-kinase) produces
lipid docking sites in the plasma membrane
• PIP3 remains in the membrane and is bound by various signaling
proteins.
 PI 3-kinase signals animal cells to survive and grow
• The TOR (target of rapamycin) protein stimulates nutrient uptake and protein
synthesis.
• Signal is terminated by phosphatases that remove the PIP3 phosphate group.
PI 3-kinase signals animal cells to survive and grow
Three classes of cell surface
receptors
 Tyrosine kinase-associated receptors depend on
cytosolic tyrosine kinase
• They depend on tyrosine phosphorylation (of their effectors) for their activity
but lack intrinsic kinase activity.
• They associate with cytosolic tyrosine kinase.
• Receptors for antigens and interleukins on lymphocytes, integrins and Src
family.
• Src family of non-receptor tyrosine kinases: Src, Yes, Fyn, Lck, Lyn, Hck, Blk
 Tyrosine kinase-associated receptors depend on
cytosolic tyrosine kinase
• They depend on tyrosine phosphorylation (on their effectors) for their
activity but lack intrinsic kinase activity.
• They associate with cytosolic tyrosine kinase.
• Receptors for antigens and interleukins on lymphocytes, integrins and Src
family.
• Src family of non-receptor tyrosine kinases: Src, Yes, Fyn, Lck, Lyn, Hck, Blk

Src:
non-receptor tyrosine kinase
Membrane bound
Has SH2, SH3 domains
Integrins associate with cytosolic tyrosine kinase
• Integrins (tyrosine kinase-associated receptor) are the main receptors the cells use
to bind to the extracellular matrix.
• Integrins binding to matrix components activate them to cluster and to form a
complex at cell periphery that allows cells to bind their substratum; the complex:
Focal adhesion.
• One of the first proteins to be recruited to this complex is the tyrosine kinase focal
adhesion kinase (FAK), followed by Src.
 Signaling by transforming growth factor-β (TGF-β)
proteins
• TGF-β act as a local mediator or hormone.
• Stimulate receptor Serine/Threonine kinases.
• Cell proliferation and death, tissue repair

1) TGF-B dimer binds serine/threonine

kinase receptor type II causing it to
associate with type I receptor and
phosphorylating it.
2) Type I serine/threonine kinase receptor
is active and phosphorylates a latent
transcription called Smad.
3) Phosphorylated active Smad
dissociate from the receptor, binds a
co-Smad protein and translocates to
the nucleus.
4) Smad/c-Smad complex binds other
proteins and activate gene
transcription.
Overlapping between downstream signaling pathways
activated by RTKs and GPCRs
 Module 5
Cellular Communication
• Cell signaling: the basics
• Cell surface receptors: G-protein coupled receptors, Enzyme coupled
receptors
• Gene expression regulation by proteolysis
Gene expression regulation by regulated
proteolysis
 Signaling between adjacent cells via Notch and
Delta regulates cell fate choices
• Lateral regulation: cells signal their neighbors to become a certain cell
type.
• Notch and Delta are single pass transmembrane proteins.
 Processing and activation of Notch by proteolytic
cleavage
• Notch undergoes 3 successive cleavages, the last 2 triggered by Delta binding.
• The cytosolic fragment translocates to the nucleus → activation of transcription.
• Once activated it cannot be used again.
Signaling between adjacent cells via Notch and Delta regulates Neuronal
cell fate
Signaling between adjacent cells via Notch and Delta regulates Neuronal
cell fate
 The Wnt / β-catenin signaling pathway
• Wnts are secreted proteins: local mediators involved in development.
• Wnt → no β-catenin degradation → it migrates to the nucleus, activates transcription
→ cell proliferation.
• 80% of colon cancers (APC).

Adenomatous Polyposis Coli

 The Wnt / β-catenin signaling pathway

• Wnts are secreted proteins: local mediators involved in development.

• Wnt → no β-catenin degradation → it migrates to the nucleus, activates transcription
→ cell proliferation.
• 80% of colon cancers (APC).
 Hedgehog signaling
• Cubitus interruptus (Ci) is normally cleaved → transcriptional repressor
 Hedgehog signaling
• Hedgehog binding to surface receptor → no Ci cleavage → transcription activation
• Skin basal cell carcinoma (Patched)
 Hedgehog signaling

• Cubitus interruptus (Ci) is normally cleaved → transcriptional repressor

• Stable Ci → Transcriptional activator
 The tumor necrosis factor-α (TNFα) / NFκB
pathway
• Stressful, inflammatory, and innate immunity responses
• Chronic inflammation and cancer
• NFκB is normally inhibited by IκB → IκB degraded upon activation → transcription
activation
 The tumor necrosis factor-α (TNFα) / NFκB pathway

• Stressful, inflammatory, and innate immunity responses

• Chronic inflammation and cancer
• NFκB is normally inhibited by IκB → IκB degraded upon activation → transcription
activation → Cell survival and inhibition of apoptosis

Degraded