9.

Cell Communication
Cells talk to each other. Lets look at two examples before
considering how this works
1. Chemotaxis individual Dictyostelium amoebae moving
towards a chemoattractant chemical
2. Quorum sensing individual bacteria that communicate
signals that can regulate light production or the ability to
take up DNA molecules.

How does the lymphocyte find the

bacterium and how does this make
the lymphocyte move?

Signals received by the front

end of the cell (these are the
soil ameba Dictyostelium)
induce actin filaments (white)
to assemble and push the cell
forward.

Here bacteria are dark until they sense that there are enough bacteria in the
flask to form a quorum sufficient to trigger them to luminesce.
To learn more about this, watch the video by Bonnie Bassler, who is making important
discoveries about quorum sensing https://www.youtube.com/watch?v=S3BOcUrJ5Vg
or read the short article by Jessica Marshall that Ill post on line.

Jessica Marshall PNAS 2013;110:2690

2013 by National Academy of Sciences

9. Cell Communication

How do individual cells find each other

and how do they known when to form
another structure?
How does a mammalian sperm find and
fertilize an egg and why doesnt it try to
fertilize every other cell along a female
oviduct?
How do cells in our body communicate
with one another during embryonic
development?
How do cells in an adult know how to
respond to changes in a body?

How do cells in our body know when to

divide and when they should not divide?

First, consider how pneumonococcus bacteria

(Streptococcus pneumoniae) can take up DNA that helps
them express genes to make them drug-resistant.

a. Low cell density

At low population density,
the concentration of the
signaling peptide is too
low to stimulate DNA
uptake by bacteria.
Receptor
Bacterial
cell
Signaling
molecule

b. High cell density

At high population density,
the concentration of the
signaling peptide is high
enough to stimulate the
DNA-uptake response.

DNA
molecule

Basic cell signaling requires a (1) signaling molecule (ligand), (2) a receptor,
and (3) molecules inside the cell that transduce the signal and make the cell
respond to the signal.
Signaling
molecule

The signaling cell

releases signaling
molecules.

Receptor

The responding cell has

receptor proteins that
bind to the signaling
molecule.

Study questions 9.1

We are familiar with cell signaling that allows cells in our bodies to communicate with other cells. Do free-living single
cells, such as bacteria or amoeba, signal each other or did signaling evolve when multicellular organisms developed?
Explain your answer.
All of the genes in a cell are simply parts of a sequence of nucleotides in a DNA molecule. Can cells incorporate DNA
from other intact or broken cells? Explain your answer and provide examples that support your answer. Your
examples may include examples from previous portions of this course.

The synthesis of cell-surface hormone receptors is regulated by the expression of genes that code for these receptors.
Assume that a receptor gene is expressed and is being translated by a ribosome. Describe the pathway by which that
receptor protein is eventually displayed on the surface of the plasma membrane in an orientation that will allow the
receptor to bind an extracellular hormone.
If all cells in a body are exposed to every hormone released into the bloodstream, why do some cells respond to a
specific hormone while other cells ignore the hormone?

Plasma membrane

Extracellular
fluid
Receptor activation

Cytoplasm
Signal transduction
Response

The signal binds to a

receptor, which is
then activated.

The signal is
transmitted to the
interior of the cell by
a signal transduction
pathway.

a. Endocrine signaling

The cell
responds, for
example, by
activating an
enzyme or
turning on
transcription of
a gene.

Termination

The response
is terminated
so that new
signals can
be received.

Signaling
molecule

Signal travels through

the circulatory system.
Receptor

b. Paracrine signaling

Plasma
membrane

Acetylcholine
binding opens
ion channels,
allowing Na+ to
flow into the
muscle cell.
Acetylcholine
Nerve cell

Muscle cell

Open

Na+
Closed

Binding site

c. Autocrine signaling

Direct contact
signaling

What can signaling do?

Regulate cell growth ex PDGF (platelet derived growth factor)
Regulate metabolism ex sugar uptake induced by insulin
Regulate embryonic development
Communicate signals from nerve cells to target cells

Unclotted
blood
Fibroblast

Clotted
blood

Plasma

Serum

Cultured with serum

Cultured with plasma

Cultured with platelet proteins

Cultured with plasma

Study questions 9.2

There are a number of different mechanisms by which cells release and receive signals. Give an example of each of
the following or describe the mechanisms by which they work:
endocrine signaling
paracrine signaling
direct contact between animal cells
direct contact between plant cells

One difficulty of stimulating mammalian cells to grow in culture is that even if you provide them with the food
they need, they do not produce a greater number of cells. Describe an experiment in which growth factors
that stimulate cell growth were identified.

You have a population of cells that respond to growth factors and will reproduce
at a reasonable pace, shown in this figure.
Now, suppose that you obtain a population of these cells that will not reproduce,
even though you add the growth factors and plenty of food.
Assuming that the growth factor stimulates cells by binding to an extracellular
receptor, propose a reason why the non-reproducing cells are no longer
responsive to the growth factors.

a. Cell-surface receptor
Polar signaling
molecule
Extracellular
domain
Transmembrane
domain
Cytoplasmic
domain
Plasma
membrane

Ligand-binding site
Activated receptor

Polar signaling
molecules cannot
cross the plasma
membrane and
rely on cellsurface receptors.

Nucleus
DNA

b. Intracellular
receptor
Small, nonpolar
signaling molecule

Activated receptorsignal
complex

Small nonpolar
signaling molecules
can freely pass
through the plasma
membrane and
activate cytoplasmic
receptors.

Intracellular receptors

Here is the structure of testosterone.

Why can it cross the plasma
membrane?

Study questions 9.3

Examine the structure of testosterone shown in the lecture slides. Explain why this would be expected to cross a
membrane rather than bind to a receptor on the external surface of a cell.
Most steroid hormones can diffuse across the plasma membrane.
a. Why might one cell in your body be responsive to a steroid hormone while another cell is not responsive?
b. Sketch a typical animal cell and describe the mechanism by which a steroid hormone can activate gene
expression. Draw the plasma membrane and any other organelles that are appropriate for your answer.

Cell signaling requires a signal molecule (ligand) and a receptor. Types of ligands that function as hormones in
humans include steroids and peptides.
a. What chemical properties distinguish steroid and peptide hormones?
b. Where are the receptors located for steroid and for peptide hormones?
Compare the pathways by which extracellular receptor is synthesized and positioned on the plasma membrane with
the pathway required for synthesis of the steroid hormone receptor.
Suppose the steroid receptor mRNA had a signal sequence to target the protein to the ER. Describe what you think
might happen to that receptor and explain your answer.

b. G protein-coupled receptor
Ligand
Receptor

G protein

GDP

GTP

Inactive

Active
Signaling
pathways
Response

c. Receptor
kinase

Phosphate
group
Response

b. Ligand-gated ion
channel
Ions

Response

Some more detailed mechanisms

Important point Well consider how a signaling molecule

(a ligand) can bind to and activate a receptor protein
but
its important to consider how a cell can stop being
activated by a signal, so think of this as we go through
some examples.

Inactive
effector

Ligand
Receptor

G protein

When the subunit is bound

to GDP, the three subunits are
joined together and the G
protein is inactive.

GDP

When the G protein is bound

by an activated receptor, the
GDP bound to the subunit is
replaced by GTP, which
activates the subunit to bind to
target proteins.

GTP
GDP

Active
effector

Second
messengers

GTP

Activated subunit binds to

and activates target protein.

Response

Inactive
adenylyl
cyclase
Adrenaline
Receptor

GDP

Activated
adenylyl
cyclase
Adrenaline
receptor

Protein
kinase A

GTP

ATP

Active
G protein

cAMP

Inactive

Active

Activated protein kinase A

phosphorylates proteins in the
heart muscle, causing heart
rate to increase.

10

Adrenaline

Activated
receptor

Each activated receptor activates multiple G proteins

which in turn activate adenylyl cyclase enzymes.
Activated
adenylyl
cyclase

Amplification

GTP

Active subunit
of G protein

ATP

GTP

cAMP

ATP
cAMP

ATP
cAMP

Each adenylyl cyclase enzyme

produces large amounts of the second
messenger cAMP, which activates
many molecules of protein kinase A.

Amplification
Inactivated protein
kinase A
Activated protein
kinase A
Amplification

GTP

ATP

Each protein kinase A

enzyme phosphorylates and
activates multiple protein
targets.

ADP

Occurs on plasma membrane

One molecule of
epinephrine binding to a
G-protein coupled
epinephrine receptor
can stimulate the
1,000,000 glycogen
phosphorylase enzymes
to release 100,000,000
glucose molecules from
glycogen.

Occurs in the cytoplasm

How cAMP signaling is turned off

GTP

Protein
kinase A
The signal molecule
adrenaline detaches
from the receptor after
a certain amount of
time, inactivating the
receptor so that it can
no longer bind to and
activate the G protein.

Activated
adenylyl
cyclase

Inactive

ATP
cAMP

Inactive

Phosphatase

Active
Active

Phosphate
group

GDP

Within a very short time, an

activated G protein deactivates
itself by converting GTP to GDP.

Phosphatases remove
phosphate groups from
proteins, causing them to
become inactive.

Phosphodiesterase
cAMP

AMP
Enzymes in the cytosol specifically
degrade cAMP which stops the
phosphorylation and activation of
target proteins by PKA.

11

Study questions 9.4

Many signaling activities in cells depend on G proteins. What distinguishes G-proteins from any other protein
associated with signaling in a cell?
G proteins can be activated when a signal ligand binds a receptor but they also are inactivated after a short time in the
cytoplasm (the time period depends on the G protein). How are G proteins activated and how are they inactivated?
Phosphodiesterase is an important enzyme associated with adenylyl cyclase signaling.
What does this enzyme do in a cell?
Choose an example of signaling associated with cAMP and describe what could happen if the cell had a mutation
that produced an inactive phosphodiesterase.
Receptor tyrosine kinases are important for many signaling activities some of which are associated with mammalian
growth and cell division. When the receptor binds a signaling molecule, the receptors dimerize (two receptors bind to each
other) and activate a tyrosine kinase.
a. What kind of molecule is a kinase?
b. What does a protein kinase do?
c. What is tyrosine?
What is the difference in function between a protein kinase and a protein phosphatase? Why are these enzymes
important in cells give one example from this lecture.
One molecule of epinephrine binding to a G-protein coupled epinephrine receptor can stimulate the 1,000,000
glycogen phosphorylase enzymes to release 100,000,000 glucose molecules from glycogen. What do glucose
and glycogen have in common and why might a cell have any reason to release glucose from glycogen (i.e.,
what use is glucose to a cell?)

Each member of the

receptor pair
attaches phosphate
groups to the other
member.

Signaling
molecules

The attached
phosphate groups
provide binding sites
for intracellular
signaling proteins.

Cytoplasmic
signaling
proteins

Phosphate
group

Inactive receptor

Dimerization

Active receptor

Signaling
molecule

Active receptor

Activated Ras
signaling protein

GTP

Inactive receptor

Active receptor

12

Inactive
kinase 1
Active
kinase 1

GTP
ATP
ADP

A small amount of signal

received by receptor
kinase is amplified when
the signal is passed
from kinase to kinase as
each is phosphorylated.

Active
kinase 2

ATP
ADP

Active
kinase 3

Changes
in gene
expression

Nucleus
Transcription regulators

Plasma
membrane

Acetylcholine
binding opens
ion channels,
allowing Na+ to
flow into the
muscle cell.
Acetylcholine
Nerve cell

Muscle cell

Open

Na+
Closed

Binding site

Study questions 9.5

Some signaling systems activate transcription.

Where, in a eukaryotic cell, does transcription occur?
What is a product of transcription?
What would a cell do with this product of transcription?
Why would it be useful for a cell to regulate transcription?

Botulism toxin contains several proteins, one of which can enter a nerve cell and prevent fusion of a synaptic
vesicle with the plasma membrane. One of these is the Botox that doctors inject into patients to prevent
wrinkles. Why does Botox treatment prevent wrinkling?

13

A gated ion channel is the

CFTR (cystic fibrosis
conductance regulator).
This channel transports
chloride ions out of a cell
and, by increasing chloride
concentration, water is
drawn out of the intestinal
cells by osmosis.
This helps keep our lungs
moist and reduces the
stickiness of mucus.

G-protein
receptor

Signal
molecule

Chloride ions
leave cell

CFTR

GDP
GTP

Adenylyl
cyclase

Phosphorylate
and activate
CFTR

cAMP
ATP

Cystic fibrosis is caused

by defects in the chloride
channel that can prevent
chloride release and the
accumulation of sticky
mucus that cannot be
removed by the cilia lining
the lungs.

Activate Protein
kinase A

Important:
Chloride is only released
when the signal molecule
activates the G-protein
receptor.

G-protein
receptor

CFTR

GDP

When the receptor is no

Adenylyl
longer activated, the Gcyclase
protein hydrolyzes GTP and
the GDP-G protein no
longer activates adenylyl
cyclase, which stops
production of cAMP

CFTR
channel
closes

GTP
cAMP
ATP

Phosphodiesterase

PkA no
longer
active

AMP

The CFTR channel

transports chloride ions out
of a cell and, by increasing
chloride concentration,
water is drawn out of the
intestinal cells and tissue
spaces by osmosis.
Chloride ions
leave cell

Cholera toxin binds to the

G-protein receptor, and
permanently activates the
G-protein.

G-protein
receptor

CFTR

GDP
GTP

Adenylyl
cyclase

cAMP
ATP

Phosphorylate
and activate
CFTR
Activate Protein
kinase A

14

So the key to G-protein

signaling is the
reversibility of the signal.

Chloride ions
leave cell

G-protein
receptor

CFTR

GDP
GTP

Adenylyl
cyclase

Phosphorylate
and activate
CFTR

cAMP
ATP

Activate Protein
kinase A

Florida

A recent cholera example

Cuba

Haiti

A Worsening Haitian Tragedy

By THE EDITORIAL BOARD, New York Times March 17, 2013
The aid group Doctors Without Borders said last Tuesday that the cholera crisis in Haiti was getting worse, for the most unnecessary
and appalling of reasons: a lack of money and basic medical supplies.
The disease has killed 8,000 people and sickened 649,000 since October 2010. International efforts to defeat the epidemic include a 10year, $2.2 billion plan for major investments in clean water, sanitation and medical infrastructure. But that is a project for the future, one
that isnt even funded yet. Doctors Without Borders says people are dying now, needlessly, because attention and money are running
out. Aid groups are leaving. Staff members at some treatment centers havent been paid in months, equipment is wearing out, and
sanitary precautions are being abandoned. The death rate has reached an intolerably high 4 percent in some places, the group said.
And the rainy season is about to make things much more difficult.
The dreadful backdrop to this emergency is an abdication of responsibility by organizations that have pledged to help Haiti, particularly
the United Nations. The U.N. said last month that it would not pay financial compensation for the epidemics victims, claiming immunity.
This is despite overwhelming evidence that the U.N. introduced the disease, which was unknown in Haiti until it suddenly appeared near
a base where U.N. peacekeepers had let sewage spill into a river.
Though the U.N. has done much good in Haiti since the 2010 earthquake, its handling of cholera is looking like a fiasco. While it insists
that it has no legal liability for cholera victims, it must not duck its moral obligations. That means mobilizing doctors and money to save
lives now, and making sure the eradication plan gets all the money and support it needs.
Its record so far is dubious. A U.N. appeal last year for $24 million for cholera programs ended the year only 32 percent financed, and in
December, the U.N. said it would contribute $23.5 million to the new 10-year plan about 1 percent of what is needed.

New York Times

March 22, 2013
Cholera in Haiti
To the Editor:
A Worsening Haitian Tragedy (editorial, March 18) points out the sad reality that cholera is
now endemic in Haiti. But it gives the impression that most aid organizations are leaving the
country at a time when thousands are dying from a preventable disease.
While some aid groups have indeed left the country or are scaling back programs, others
have made fighting the epidemic their top priority.
Since the cholera outbreak began in 2010, AmeriCares alone has delivered enough
medicines to treat about 150,000 patients, and we continue to expand our Haitian staff to
ensure that we can continue stocking cholera treatment centers all across the country with
medicines and supplies.
Going forward, N.G.O.s in Haiti must come together and intensify focus on prevention
activities that offer the best hope for preventing needless deaths, unnecessary
hospitalizations and unquantifiable suffering. Better coordination and leadership will bring
about more lasting change than any amount of recrimination and finger pointing.

No one should have to die from a disease that can

be avoided with soap and water, and the collective
will to make them available.
CURT WELLING
President and Chief Exec., AmeriCares
Stamford, Conn., March 19, 2013

15

So CFTR channels open because

A signal
binds to G-protein receptors that
Activate G-proteins by exchanging GDP for GTP which
Binds to and activates adenylyl cyclase which
Converts ATP to cyclic AMP which
Binds to and activates protein kinase A which
Phosphorylates and opens chloride channels and
Chloride ions draws water from tissue spaces and
Hydrates mucus

So CFTR channels close

When no more water release is needed, the signal releases
from the G-protein receptor
G proteins hydrolyze GTP and become inactive GDP-G proteins
Adenylyl cyclase stops working and cAMP is converted to AMP
by phosphodiesterases
Without cAMP, the protein kinase is no longer active
Without active PkA the CFTR channel closes.

The cholera toxin modifies the G protein, which

keeps the G protein in the GTP-G protein stage

Adenylyl cyclase keeps working and produces cAMP is

cAMP stimulates and keeps the protein kinase active
This keeps the CFTR channels open
Water is drawn from tissues and into the intestine
Which results in diarrhea, water loss, and possible
death

16

Study questions 9.6

Cholera toxin, released from Vibrio cholerae bacteria, induces diarrhea and dehydration. Infected individuals die
because they are dehydrated, due to water loss from the bloodstream. Water is lost because chloride channels that
bind and are activated by cyclic AMP release chloride ions into the intestine. This increase in salt (chloride and
sodium ions) draws water (by osmosis) out of the bloodstream and into the intestine, resulting in diarrhea. Cholera
toxin acts on the G-protein and prevents GTP hydrolysis.
a. Sketch the process by which the binding of a bacterial protein opens chloride channels. (Hint chloride
channels are ligand-gated ion channels).
b. Cholera toxin blocks GTP hydrolysis by the G protein. Why does it induce diarrhea?
c. Based on your knowledge of osmosis, how can osmosis induce movement of water from tissue into the
intestine?
d. How can infection of a human with Vibrio cholera cause diarrhea?
d. Without proposing antibiotics, to kill bacteria, or miracle drugs, to block the action of cholera toxin, propose a
simple and inexpensive method that might be (is) used to treat patients infected with Vibrio cholerae bacteria.

Study questions 9.7

Receptor tyrosine kinases are important for many signaling activities some of which are associated with mammalian
growth and cell division. When the receptor binds a signaling molecule, the receptors dimerize (two receptors bind to each
other) and activate a tyrosine kinase.
a. What kind of molecule is a kinase?
b. What does a protein kinase do?
c. What is tyrosine?
Compare the functions of protein kinases and protein phosphatases. Give one example from this lecture to
illustrate their importance in cells..
One of the key properties of signal transduction pathways is the ability of each signaling molecule in the pathway to be
turned on or turned off.
Consider each of the following pathways and identify, for each step in the pathway,
(a) the components that are turned on,
(b) the mechanism or molecule that activates each molecule,
(c) the mechanism or molecule that inactivates the activated molecule
(d) what would happen to the process if any one of the steps were not able to be inactivated.
Pathway:
1. A pathway in which a ligand stimulates a G-protein coupled receptor to open a CFTR (cystic fibrosis
channel protein) channel to allow chloride ions to diffuse out of a cell. Your answer should include the
mechanism by which the CFTR channel is opened (within the limits of what we discussed in lecture), the
mechanism by which the CFTR channel would be closed, and what would happen if GTP could not be
hydrolyzed by G-proteins associated with this process. (This question and answer should be similar to the first
question in this set)
2. A pathway by which the binding of a single epinephrine molecule to a cell-surface G-protein receptor can
induce the release of 100,000,000 glucose molecules from stored glycogen molecules. Your answer should
include the steps by which the production of glucose from glycogen occurs and what happens if epinephrine
is no longer bound to the receptor.

Types of G proteins
There are two major classes of G proteins
Monomeric G proteins
Heterotrimeric G proteins
What is a difference between a monomeric and a
heterotrimeric G protein?

All are inactive with GDP bound and active with GTP bound

17

Heterotrimeric G proteins
Composed of 3 protein
subunits - , ,

These are coupled to Gprotein receptors we have

discussed in class

When the receptor is activated, GTP is bound to G and the

G dissociates from G and G.
The G and G then can target different target molecules

Monomeric G proteins (small G proteins)

More than 100 different small G proteins that can be grouped
in 5 classes:
Ras

Rho

Rab

Ran

Arf

Have a variety of functions in membrane trafficking and other

aspects of cell regulation

Importance: mutations in Ras alone are found in ~25% of all

human tumors. Ras is particularly important in signaling
pathways that stimulate cell growth or inhibit cell death
(apoptosis).

18

Scaffold proteins can bring several signaling

components together.

Study questions 9.8

Scaffold proteins are found in a number of cells, With regard to signaling, what might be a function of a
scaffolding protein?

19