Review Session for Exam 3

topics to cover
EGF vs. NGF endosomal signaling
anchoring & scaffolding proteins
signaling crosstalk
post-transcriptional regulation
transcriptional control
CRE/CREB, kinase signaling pathways
nuclear receptor signaling
chromatin structure
epigenetic regulation
EGF vs. NGF signaling
EGF vs. NGF signaling
 EGF vs. NGF signaling

EGFR
 EGF vs. NGF signaling

GFR: clathrin-mediated endocytosis

EGF vs. NGF signaling

Trk
 EGF vs. NGF signaling
Trk: clathrin-independent endocytosis

black dots = EGFR

arrows =
clathrin-coated pit

** Trk is
black dots = TrkB endocytosed
in a manner
that is
independent
of clathrin
 EGF vs. NGF signaling
Trk: Pincher-mediated endocytosis

control:
normal ** Trk endocytosis is Pincher-
endocytosis dependent
 EGF vs. NGF signaling
Trk: Rac-mediated endocytosis

control:
normal ** Trk endocytosis is Rac-
endocytosis dependent
EGF vs. NGF signaling
Trk: Pincher/Rac-
mediated endocytosis
mechanism

x x

x x
 EGF vs. NGF signaling
molecular basis: endocytosis

Trk endosomes avoid Ra

EGF vs. NGF signaling
retrograde endosomal signaling in the
axon
retrograde endosomal signaling in the
axon
retrograde endosomal signaling in the
axon
retrograde endosomal signaling in the
axon
the chamber culture model
retrograde endosomal signaling in the
axon
the chamber culture model
retrograde endosomal signaling in the
axon
the chamber culture model

multi-vesicular body
retrograde endosomal signaling in the
axon
the chamber culture model

are they
active?
retrograde endosomal signaling in the
axon
the chamber culture model
retrograde endosomal signaling in the
axon
the chamber culture model
retrograde endosomal signaling in the
axon
molecular components of
the signaling endosome
 anchoring & scaffolding proteins

nomenclature:
anchoring:
bringing signaling components to a specific
subcellular location
scaffolding:
bringing multiple signaling components
together in close proximity to each other
purpose?
efficient signaling
anchoring & scaffolding proteins
anchoring & scaffolding proteins
 anchoring & scaffolding proteins

ARMS
 anchoring & scaffolding proteins

Gab1
 anchoring & scaffolding proteins

Par polarity complex

 anchoring & scaffolding proteins

AKAP (A-kinase anchoring protein)

green = PKA
 anchoring & scaffolding proteins

AKAP (A-kinase anchoring protein)

 anchoring & scaffolding proteins

AKAP (A-kinase anchoring protein)

 anchoring & scaffolding proteins
post-synaptic density
 anchoring & scaffolding proteins
post-synaptic density & PDZ domains
anchoring & scaffolding proteins
convergence on adenylyl cyclase
convergence on PLC
Ca2+ crosstalk

PI3K

PKB/Akt
endpoint convergence: TH
endpoint convergence: CREB
computational modelig
control of gene expression
RNA interference
 post-transcriptional
regulation
translational
regulation

Erk
MNK1
eIF4E
 post-transcriptional
regulation
translational mTOR
regulation eIF4E
4E-BP
 Thestepsofeukaryoticposttranscriptionalprocessing

Decay / storage
Transcription
P-bodies / Stress Granules

1 2 3 3 4 5 General
translation
Local AAA
A
translation
AA
splicing 1 2 3 4 5 12345

Cleavage/ 1 2 3 4 5 A200
polyadenylation

mRNP
packaging

Nucleus Export

Cytoplasmic
factors mRNAtransport/localization
Localized mRNAs play a critical role in the local control of translation
Induced localization and translation of mRNAs
control axon growth
transcriptional control
 transcriptional control:
cAMP responding element (CRE)
1986

necessary &
SST 5 flanking sufficient for
cAMP-mediated
response

palindromic
CAT sequence
(TGACGTCA)
within 100 bp of
TATA-box

less active if
moved further
 Common Reporter Assays

Chloramphenicol Acetyltransferase Reporter Lucipherase Reporter

increasing
CAT
acetylated

unacetylated

chromatography
 transcriptional control:
CRE-binding protein (CREB)
Protein Kinase A acts in the nucleus Enhancers are binding sites for activators
 transcriptional control:
CRE-binding protein (CREB)
Alpha-helix

MG 2+
 transcriptional control:
CRE-binding protein (CREB)

MAPK

rsk
 transcriptional control:
CREBs interaction partner
transcriptional control:
JAK/STAT signaling
 transcriptional control:
serine/threonine kinase receptors
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
chromatin structure remodeling
 transcriptional control:
chromatin structure remodeling
 transcriptional control:
chromatin structure remodeling
 transcriptional control:
chromatin structure remodeling
the histone code
 transcriptional control:
chromatin structure remodeling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling

on

off
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
hormone & nuclear receptor signaling
 transcriptional control:
chromatin structure remodeling
transcriptional control:
epigenetic modulation
transcriptional control:
epigenetic modulation
transcriptional control:
epigenetic modulation
 transcriptional control:
epigenetic modulation

secondary response genes

primary response genes

(NGFI-A)
 transcriptional control:
epigenetic modulation

GR levels

?
behavior

high GR levels low GR levels

 transcriptional control:
epigenetic modulation

GR

low stress behavior high stress behavior

transcriptional control:
epigenetic modulation
 transcriptional control:
epigenetic modulation

GR levels

behavior
 transcriptional control:
epigenetic modulation

behavior

GR levels
 transcriptional control:
epigenetic modulation

behavior

GR levels