View
227
Download
0
Category
Tags:
Preview:
Citation preview
SignalTransduction
Pathways
Signaling Pathways ControlMany Cellular Processes
Cell CycleCell PolarityCell Shape
Development ….. and DiseaseDevelopment ….. and Disease
Signals
Stop!
Go Faster?
Go.
……….Pathway OFF
….…Basal
……..Pathway ON
Stimulus
Receptor
PAK/G-protein
MAP KinaseCascade
Txn Factors
Generic MAP Kinase Pathway
Cell BiologyGFP Collection
GenomicsDNA Microarrays
Yeast GeneticsGenomic Screens
BiochemistryTAP Tag Collection
Genetics and High-throughput Technologies
Budding Yeast is a Model Organism
Filamentous Growth (FG) in Yeast
FG is required for fungal FG is required for fungal pathogenesis.pathogenesis.
Saccharomyces cerevisiae
PRP
Ste2/3/4/18
Ste20/Cdc42
Ste11Ste7Fus3
Ste12
Ste5
FGP
Sho1 + ?
Ste20/Cdc42
Ste11Ste7Kss1
Ste12/Tec1
HOGP
Sho1
Ste20/Cdc42
Ste11Pbs2Hog1
Hot1
FGP
?
Ste20/Cdc42
Ste11Ste7Kss1
Ste12/Tec1
MAPK Pathways Share Factors
Mutants defective for agar-invasion (FG)
Genome-Wide Genetic Screen
UP
DOWN
NO CHANGE
DNA Microarray Analysis
Comparison of Genomic Data
MSB2 was identified in both screens.
Scaffold at the Head of the FG Pathway
Msb2
Cdc42 Sho1
Mucin Domain
Role of Mucin Domain in Signaling
Mucin domain is inhibitory.Mucin domain is inhibitory.
MSB2 Δmucin
1220swt wt
50
10
40
30
20
FU
S1-l
acZ
N…(698)S Q V S D T SV S Y T T S S S S V S Q V S D T PV S Y T T S S S S V S Q V S D T PV S Y T T S S S S V S Q V S D T PV S Y T T S S S S V S Q V S D T PV S Y T T S S S S V S Q V S D T SV P S T S S S S S V S Q V S D T PV P S T S S S S S V S Q (818) …C
Go, Go, Go!
Hyperactivity
Questions about the FG pathway
What is Msb2 sensing?
How are general factors specified by Msb2?
In what other ways is specificity achieved?
Stop!
Negative Regulation
FGP
Sho1 + Msb2
Ste20/Cdc42
Ste11Ste7Kss1
Ste12/Tec1
HOGP
Sho1
Ste20/Cdc42
Ste11Pbs2Hog1
Hot1Rck2
Novel Negative Regulator: a Calmodulin Kinase
Questions about Rck2
Where does the kinase act?
How does P~lation by Rck2 cause pathway shut-off?
Is the response specific?
Rotate in Paul Cullen’s Lab?
Sure!
Training in Paul Cullen’s Lab
Question-based approach
Learn many different techniques (Genomics)
Working with yeast is Fun!
Recommended