Signal Transduction

Review cell-cell junctions (ch11.1-2) for

structure-function relationships (see
slides) as they relate to tissue function
You dont need to know them for exam
4
We will revisit them later in Unit 5
2

Would you like to attend 3-4 short

student talks for clicker credit?
If yes, which time?
A.
B.
C.
D.
E.

Monday 10 10:50 AM
Monday 11 - 11:50 AM
Monday 4 - 4:50 PM
Wednesday 1 1:50 PM
Wednesday 2 - 2:50 PM

Talks will be given

week of 11/30 and 12/7
Limited to 25
students per time slot
If sufficient interest,
registration will be
online through
courseweb

3 points for attending talk, +2 more if you ask a relevant question

Students will be able to describe how cells

transduce extracellular signals into internal
events, such as turning on genes, entering
mitosis, or altering metabolism.
transform a signal from one medium or form to another

Focus:

Lipid signals and receptors

Signal transduction cascades

Initial transduction event

Amplification

Signaling through Hormones

Local:
Factors

Distance
Factors

secreted into extracellular matrix

(systemic):
secreted into bloodstream

Challenge: can you think of examples where a local signal

is preferable to a whole-body signal, and vice versa?
Example: histamine
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Receptor proteins resemble enzymes

SUBSTRATES

Selective
active site

LIGAND

Shape
change
Effect

Receptor proteins resemble enzymes

Variable reaction
rates
Regulated by:
phosphorylation
allosteric
deactivation

CATALYZATION

TRANSDUCTION
7

Ligand lookalikes have two effects:

ANTAGONIST
X

AGONIST
+

X
BLOCKS
signal transduction

ACTIVATES
signal transduction
without ligand
8

General rule #1
Response to a signal is dependent
upon CELL TYPE
receptors

(if any)
signal transduction elements inside?
Function/specialization of cell?
9

In a multicellular organism, different

cells types make different receptors.

(Differential gene expression = different proteins present)

10

EFFECT of a signal depends upon:

(1) does the cell receive the signal?
(2) does the cell have a receptor for the signal?
A and B exposed,
only A responds

A.

B.

C.

Local signals: diffuse through ECM

D.

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EFFECT of a signal depends upon:

(1) does the cell receive the signal?
(2) does the cell have a receptor for the signal?
All cells exposed,
only A and C respond

A.

B.

C.

Systemic Signals travel through bloodstream

D.

12

EFFECT of a signal depends upon:

(1) what does the receptor do when it receives the signal?
(1) initiates signal transduction
(2) ultimate effect dependent on cells specialization

A.

B.

C.

D.

opens an ion channel

inhibits an enzyme

turns a gene on or off

turns on an ion pump

13

EFFECT of a signal depends upon:

A.
opens an ion channel
turns on a gene

B.
If

C.

D.

is released into the bloodstream,

which cells will respond to it?

turns off a gene

inhibits a pump protein

A. all of them
B. A and B

Ligand for any

green receptor

C. C and D
D. A and C
E. none of them

14

EFFECT of a signal depends upon:

A.
opens an ion channel
turns on a gene

B.

C.

D.

Will cells
If
is released
A and C have
into the
thebloodstream,
same
which cellstowill respond
response
?
to it?

turns off a gene

inhibits a pump protein

A. yes
all of them
B. no.
A and B

Ligand for any

green receptor

C. C and D
D. A and C
E. none of them

15

EFFECT of a signal depends upon:

x
x

A.

B.

C.

D.

x
opens an ion channel

We mutate the gene that encodes

turns on a gene
turns off a gene

What changes when

blood?

inhibits a pump protein

A. no cells can respond

is present in

B. A can no longer respond, but C can

C. C can no longer respond, but A can
D. nothing- cells are normal
16

General rule #2:

lipid signals are not transduced
Receptors in the cytoplasm
Generally turn on genes.
Challenge: describe in your own words what properties of molecules
determine whether they can freely cross the cell membrane
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This is NOT transduction

Reception:
cytoplasmic
Response:
turn on genes
OR alter internal
membrane
properties
18

Example: the androgen receptor

is also a transcription factor
DNA binding
domain
N-

-C
send me to the nucleus to
turn on MANLY genes!

Nuclear Import signal

(blue)

Send me back to the

cytoplasm

Nuclear Export signal

(red)
(much stronger)
19

Example: the androgen receptor

is also a transcription factor
DNA binding
domain
N-

androgen

-C

send me to the nucleus to

turn on MANLY genes!

Steroid hormone covers and blocks the export signal

20

General rule #3: If a signal cannot cross the

membrane by itself, it is transduced.
Signal
binds
receptor

Receptor activates
cellular chain reaction

The signal itself

never enters the cell.

TRANSDUCTION
Two types of transduction:

21

Transduced
signals are
amplified
two types of amplification:

Cell response (variable):

Open channels
Turn on metabolic pathways
Turn on genes
22

Two types of initial signal transduction

extracellular signal intracellular signal

G-protein

coupled receptors

Activate

G-proteins
G-proteins activate next step
Enzyme-linked
Directly

receptors

activate enzymes via a protein bridge

KNOW the two kinds of receptors and how they transduce

23

Two types of signal amplification

Both types can coexist in the same pathway
Second

messengers: small, nonprotein

signal molecules
DAG
Ca2+
IP3
cGMP

Phosphorylation

cAMP

cascades

a series of protein kinases activating each other

KNOW the two kinds of amplification discussed in this lecture

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Many variations of signal

transduction
GPCR

Second messengers

Effect 1
Effect 2

Enzymelinked

Phosphorylation
cascade

Effect 3

Main point: we will cover two examples. There are many

other possible combinations in different cells
25

G-protein coupled receptors share

common features
one subunit
7 transmembrane
domains
heterotrimeric
G-Protein binds
, ,

26
http://en.wikipedia.org/wiki/G_protein-coupled_receptor

Common G-protein-coupled
receptor response:
1. activate enzyme

Phospholipase C (PLC)
or Adenylyl cyclase (AC)

2. enzyme makes second messenger

3. messenger triggers response
27

Common GPCR second messengers:

The PIP2 signaling pathway

Enzyme: PLC
Second messengers:
IP3, DAG, and Ca2+

PLC is activated by G
proteins.
PLC cuts PIP2 into IP3 and
DAG
IP3 opens Ca2+ channels
Ca2+ released into cytoplasm

DAG + Ca2+ activates PKC

PKC phosphorylates other
proteins
28

Common GPCR second messengers:

The cAMP signaling pathway

Enzyme: AC
Second messenger: cAMP

Adenylyl cyclase
Extracellular
Intracellular

AC is activated by G
proteins.

ATP

AC turns ATP into cAMP

cAMP
Open ion
channels

PKA

Phosphorylation
of substrates

cAMP mainly activates

protein kinase A
PKA phosphorylates
other proteins
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Two types of initial signal transduction

extracellular signal intracellular signal

G-protein

coupled receptors

Send

activated G-proteins floating off to

activate enzymes

Enzyme-linked
Directly

receptors

activate enzymes via a protein bridge

30

Receptor tyrosine kinases are

single subunits that dimerize when
ligand binds

http://membranereceptors.com/Test2/wp-content/uploads/2013/07/Enzyme-linkedreceptors1.jpg

Common enzyme-linked receptor

signaling pathway: activate Ras and Raf

Signal received

Autophosphorylation:
receptor
phosphorylates itself

this is not a
G-protein

AMPLIFICATION at every step

Receptor activates
Ras

(RAF)

Ras
phosphorylates
Raf
Raf
phosphorylates
another kinase
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Example: MAPK (mitogen-activated

protein kinase) signal cascade
Mitogen: something that triggers a cell to divide (initiate mitosis)

RAS is activated
Bridging proteins activated

GTP
GDP

mitogen
EX: Epidermal growth
factor (EGF)

SOS
GRB2

RAS

EGF
receptor P

Tyrosine inactive
kinase activity
activated

P
RAF

P
MEK

P
MAPK

RAF is activated MEK is activated

MAPK is activated

Transcrip. Factor
Transcription factortranscription
moves tofactor
activated
nucleus to turn on genes
NOTE: GTP binding protein =/= G protein

33

Representing signal transduction

pathways
A

Inhibition is represented
with a bar

Inhibitor
Arrows indicate a protein
directly activating another
protein

often the mechanism of activation is

phosphorylation (not shown)

these are not metabolic pathways

elements in circles
proteins or second messengers

the components
(A,B,C,D) ARE the
enzymes

34

Heres an example from literature:

Challenge: identify the

following components:
1. a G-protein coupled
receptor, and which
enzyme it activates
2. an enzyme-linked
receptor, and the
second messenger
pathway it activates
3. an inhibitor, and what
it inhibits.

Miyashiro K , Eberwine J. PNAS 2004;101:17329-17330

35

Crosstalk

Enables cell to
integrate signals

Reaction to B
different than
reaction to A + B

36

Major intracellular signaling pathways mediated by small GTP-binding proteins. Three different MAPK signaling pathways named ERK pathway
(Ras
Raf
MEK1/2
ERK1/2), SAPK/JNK pathway (MEKK1
SEK/MKK4
SAPK/JNK), and p38 pathway (TAK1
MKK3/6
p38) are well
established and involved in the intracellular signaling of the small GTP-binding proteins. From these MAPK pathways, various transcription factors such
as Elk1, ATF-2 (CREB), AP-1, NF- B, and Sp1 are activated and start the transcription of target genes.
From Kim et al (2000) Effect of lovastatin on small GTP binding proteins and on TGF- 1 and fibronectin expression. Kidney International 58, S88S92

37

You should be able to:

When given a signal, determine where receptor

will be (intra- or extracellular?)

Know the three types of receptors discussed here

(there are many others but you will learn them in
other courses)
Which

act directly on genes

Which ones stay in the membrane and transduce
signals
What transduction means
What agonist and antagonist mean
38

You should be able to:

Understand signal amplification

Why cytoplasmic receptors (like androgen receptor example) are NOT

amplified
How phosphorylation cascades work
How second messengers work
How phosphorylation cascades and second messengers amplify signals

Be able to interpret a simple signal transduction diagram

Easily confused with metabolic pathway diagrams

Know the inhibits symbol
Be able to identify the two receptors and identify the types of signal
transduction in the diagrams

39

You should be able to:

On the next exam you will get a new signal transduction pathway
ahead of time. You will also be given signal transduction pathways
that are cancer, immunity, and development-related. You should be
able to:
Identify signals and receptors, even if signals/receptors are cellular
adhesion proteins or other proteins that have other functions
Identify the events that occur in the cell when the signal is received,
including ultimate effect on the cell itself
Predict what will change if an element of the system is mutated,
blocked, or otherwise altered, including:

Mutating a signal or receptor

Blocking a receptor with an antagonist
Using a receptor agonist
Mutating a kinase so that it is always ON

See next slide for clarifications about amplification

40

Clarification/elaboration of
some concepts covered in
signal transduction
(1)
(2)
(3)

Naming conventions
Signal amplification
Signal deactivation
41

Naming conventions
A phosphorylates B.

A is a B Kinase

A
B phosphorylates C.

B is a C Kinase

42

Naming conventions
A

1. D is (for example) MAP kinase

Abbreviated MAPK

43

Naming conventions
A

2. C is then D kinase
AKA MAPK kinase

abbreviated MAPKK
D

1. D is (for example) MAP kinase

Abbreviated MAPK

44

Naming conventions
3. B is then C kinase
AKA MAPKK kinase

abbreviated MAPKKK
B

2. C is then D kinase
AKA MAPK kinase

abbreviated MAPKK
D

1. D is (for example) MAP kinase

Abbreviated MAPK

45

Signal amplification

46

Signal amplification
inactive

Activation
through addition
of a P group

active

inactive

Deactivation
through removal
of a P group

47

Signal amplification
inactive

Activation
through addition
of a P group

active

inactive

Deactivation
through removal
of a P group

While active, it can catalyze reactions

(whatever they are)

48

Signal amplification
inactive

Activation
through addition
of a P group

active

inactive

Deactivation
through removal
of a P group

If this enzyme is a kinase,

it will phosphorylate its substrates.

49

Signal amplification
inactive

Activation
through addition
of a P group

active

inactive

Deactivation
through removal
of a P group

P
P
It will catalyze more than one reaction as long as
it is active

P
P

P
P

P
50

Signal deactivation

51

Signal deactivation
inactive

Activation
through addition
of a P group
Enzymes that add P
groups are called
kinases

active

inactive

Deactivation
through removal
of a P group
Enzymes that remove P
groups are called
phosphetases

52

Signal deactivation
kinases

P
active

inactive
phosphetases

For every on signal there is an off signal

53

Signal deactivation
kinases
phosphetases

No signal: all proteins are in the off state

54

Signal deactivation
kinases
phosphetases

Signal: Kinase is activated.

Proteins are phosphorylated and enter the active
on state

55

Signal deactivation
kinases
phosphetases

Signal: Kinase is activated.

Proteins are phosphorylated and enter the active
on state
BUT proteins are also being dephosphorylated
(deactivated) by active phosphetases
56

Signal deactivation
kinases
phosphetases

AS LONG AS THE SIGNAL is active

the kinase will be active
and proteins DEACTIVATED by
phosphetases
will be REACTIVATED by the kinase
57

Signal deactivation
kinases
phosphetases

As soon as the signal turns off

58

Signal deactivation
kinases
phosphetases

As soon as the signal turns off

Phosphetases rapidly deactivate any remaining
active proteins. Signal is halted.

59