Wnt Signalling and Cancer Flashcards
How many cells does the human body consist of?
50-100 E12
What happens at the membrane during signal transduction?
Receptor/ligand interaction
What happens in the cytoplasm during signal transduction?
Transduction of the signal
What happens in the nucleus during signal transduction?
Transcription is switched on/off
What is cancer a disease of?
Deregulated signal transduction leading to uncontrolled proliferation
Wnt = Wg + Int-1. What do Wg and Int-1 represent?
Wg is drosohila segment polarity gene and Int-1 is the mouse proto-oncogene
In evolution, when did the Wnt pathway appear?
At least 650 million years ago
What are the two types of Wnt pathway?
Canonical and non-canonical
In canonical Wnt signalling, what happens at the cell membrane?
Wnt ligands bind receptors Fz and LRP-5 and LRP-6
What does Fz stand for?
Frizzled
What does LRP stand for?
Low density lipoprotein receptor-related protein family
In canonical Wnt signalling, what happens in the cytoplasm?
Translocation of axin to the membrane and binding to the intracellular tail of LRP
Phosphorylation of Dsh (Dvl)
Accumulation and nuclear translocation of beta-catenin
What does Dsh and Dvl stand for?
Dishevelled
In canonical Wnt signalling, what happens in the nucleus?
New interactions involving TCF/LEF proteins
Transcriptional repression/transcriptional activation switch
Describe the Wnt family of ligands
Consists of 19 glycoproteins which are rich in cysteine
Wnt ligands can be divided into tow groups according to their ability to transform what?
Mouse epithelial cell line C57MG
What are the transforming Wnt ligands?
1, 3, 3a and 7a
What are the non-transforming or weakly transforming Wnt ligands?
2, 4, 5a, 5b, 6, 7b, 11
All Wnt ligands are palmitoylated, what does this mean?
They have a covalent attachment of fatty acids making them hydrophobic and poorly soluble in water
What does CRD stand for?
Cysteine-rich domain
What family is the Wnt receptor frizzled a member of?
The seven-transmembrane-spanning (7TMS) superfamily
What was the first Wnt receptor to be identified?
Frizzled
How was frizzled first identified?
Through genetic studies in drosophila melanogaster
Describe frizzled proteins
Encoded by about 10 conserved genes in humans and other mammals Are a small divergent subclass of 7TMS within the G protein-coupled receptor superfamily
What are LRPs specifically required for?
Beta-catenin (canonical) signalling
What is required for the association of LRP with Fz?
Wnt
What was a result of enforced association when LRP6 was fused with the neurotrophic receptor and Fz with its ligand NT3?
Wnt-independent beta-catenin signalling
Give an example of a Wnt agonist
R-spondin
What does R-spondin do?
Binds LGR4/5/6 resulting in internalisation and degradation of RNF43 and ZNRF3 E3 ligases
What do ZNRF3 E3 ligases do?
Ubiquitinate Fz
What does LGR stand for?
Leucine-rich repeat containing G protein coupled receptors
Give an example of a Wnt antagonist
Dickkopf
What does dickkopf (Dkk) do?
Prevents signalling by ninding LRP5/6
What is Kremen-2 involved in?
Internalisation and degradation
What is beta-catenin called in drosophila?
Armadillo
What is each armadillo repeat in beta-catenin composed of?
A pair of alpha helices that form a hairpin structure
Multiple copies of the repeat form an alpha-solenoid structure
What do beta-TrCP proteins ensure?
Ubiquitination and proteasomal degradation of beta-catenin
What does beta-TrCP stand for?
Beta-transducin repeats-containing proteins
Where does Dvl act in the Wnt pathway?
Immediately downstream
What do the proteins that interact with Dvl define?
Whether canonical or non-canonical pathways are being activated
Which Wnt pathway does phosphorylation of Dvl by casein kinase I (CKI) promote?
The canonical pathway
Is axin an activator or inhibitor of the Wnt pathway?
Inhibitor
What are the immediate consequences of ligand-receptor interaction?
Dvl phosphorylation, Dvl/Fz, and LRP/axin interaction
How is the destruction complex inactivated?
Using a two-signal (two branches) model of Fz-LRP signalling to beta-catenin
Two branches: Fz/Dvl and LRP/axin talk through axin-Dvl interaction
What do axin and APC form and what does this allow?
Forms a structural scaffold
Allows GSK-3beta to phosphorylate beta-catenin and axin
What is the axin/APc complex able to recruit and what does this allow?
CK1 which phosphorylates beta-catenin promoting its phosphorylation by GSK-3beta
What is the axin/APC complex central to?
The regulation of beta-catenin levels in the cytoplasm of non-stimulated cells
What does FAP stand for?
Familial adenomatous polyposis
What is the biggest part of the scaffold that is found mutated in FAP and sporadic colorectal cancers?
APC
Where is GSK3beta active?
In unstimulated resting cells
What does GSK3beta stand for?
Glycogen synthase kinase 3beta
What does phosphorylation by GSK3beta usually inhibit?
Substrates such as beta-catenin
What is GSK3beta?
A serine/threonine kinase
What does GSK3beta have a preference for?
Substrates with a pre-existing phospho-ser-thr, four residues C-terminal of the ser/thr site to be phosphorylated
What is the role of CK1alpha?
Primes beta-catenin for phosphorylation by GSK3beta
What is the role of diversin?
Recruites CK1alpha to the degradation complex
How many Tcfs are there in mammals?
Four
The beta-catenin/armadillo-binding domain and the DNA-binding HMG box are best conserved, true or false?
True
Are the C-termini of Tcfs conserved or unique?
Unique to one or a few members
How many alternative C-termini does splicing result in for hTCF-1 and -4?
Two
What does alternative promoter usage, as described for hTCF-1 result in?
The absence of the beta-catenin binding domain
What does HMG stand for?
High mobility group box
What is HMG?
A conserved domain responsible for DNA binding
Interacts with the (A/T)(A/T)CAA(A/T)GG DNA motif
What is pangolin?
A tcf in drosophila
What screen can be carried out to analyse the proteins that interact with Tcfs?
Yeast two-hybrid screens