Lecture 16 & 17 (TSGs) Flashcards
2 best studied tumour suppressor proteins
Rb and p53
What happens when tumour suppressor proteins are implicated in cancer?
They have lost their function - no longer able to negatively control cellular growth
Many tumourigenic DNA viral oncoproteins sequester…
Rb and p53
Tumorigenic DNA viral oncoproteins that interact with Rb (under-phosphorylated)
- SV40 large T
- Adenovirus E1A
- HPV E7 antigen
Tumorigenic DNA viral oncoproteins that interact with p53
- SV40 large T
- Adenovirus E1B55K
- HPV E6 antigen
- EBNA-1
How does Rb negatively control cellular growth?
- by repressing expression of genes that stimulate cell proliferation through its ability to bind E2F transcription factors & inhibit their activity
- by activating expression of genes that inhibit proliferation
Two Rb-related proteins
p107 and p130 (pocket proteins)
Where do most mutations which inactive Rb occur?
in its pocket region
Rb pocket domains shown strong homology to which 2 general transcription factors?
TBP and TFIIB
How can transcriptional regulation by Rb be mediated?
- E2F (transcriptional activator)
- ATF (nuclear oncoprotein)
- RCE (retinoblastoma control element) sites
What transcription factor is implicated in Rb control of transcription from RCE sites?
SP-1
How does Rb function in terminal differentiation of muscle cells?
by preventing binding of MyoD dimers to DNA
The overall structure of the Rb protein depends on its __ state
phosphorylation
Most frequently implicated gene in human cancers
p53 (found inactivated - deleted/mutated)
p53 is a nuclear transcription factor that’s particularly active in response to…
cellular stress
Normal function of p53
To sense if problems are occurring within the cell (e.g. if there’s extensive nucleic acid damage, p53 will induce expression of genes involved in DNA repair and if this is unsuccessful, p53 will induce expression of genes involved in apoptosis)
p53 functions as a __ and is subject to __
tetramer
phosphorylation
~50% of ovarian cancer tumours have a __ mutation
p53
Motif for p53 binding
5’ Pu-Pu-Pu-C-(A/T)-(A/T)-G-Py-Py-Py 3’
4 domains of p53
- N-terminal transactivation domain
- DNA-binding domain (contains a zinc ion)
- Oligomerisation domain
- C-terminus regulatory domain
How many different promoter regions can p53 bind?
~300
p53 is normally present in __ levels in cells
low
p53 is typically bound to __ , which regulates its turnover
MDM2
Examples of events that will lead to the activation of p53
- lack of nucleotides
- UV radiation
- ionising radiation
- oncogene signalling
- hypoxia
- metabolic stress
- blockage of transcription
How does MDM2 regulate p53 turnover?
MDM2 expression is controlled by p53 (auto-regulatory loop). When MDM2 is expressed, it will associate with p53 and flag it for degradation. If problems arise within the cell, MDM2 will separate from p53 to liberate p53 activity.
What is MDM2?
a ubiquitin ligase
Activation of which kinases (due to DNA damage) leads to p53 phosphorylation, preventing it from binding to MDM2?
ATM and DNA-PK
Examples of stress-activated kinases that perturb p53/MDM2 association
- ATR
- Casein kinase II
Activated oncogenes such as Ras can induce the activity of another tumour suppressor protein called __ , which can bind MDM2
p14 ARF
3 ways to prevent p53 degradation by MDM2
- activated kinases
- activated oncogenes
- auto-regulatory loop
p53 elicits cell cycle arrest in response to DNA damage through transcriptional induction of a gene called __
p21
What does p21 inhibit?
several cyclin-CDK complexes
p53, p21 and __ act together to sense DNA damage, halt cell cycle and repair DNA
PCNA
What is NF-1?
a GTPase activating protein (GAP) for Ras
NF-1 is __ when it’s implicated in cancer (neurofibromatosis)
inactivated (unable to stimulate Ras to hydrolyse GTP)
How can overexpression of Rev-1 reverse Ras-induced transformation?
believed to do so by abortively sequestering Ras effector proteins like Raf and PI3K
Genes implicated in hereditary breast and ovarian cancer (HBOC) syndrome
BRCA1 and BRCA2
Tumour suppressor protein implicated in many colorectal cancer cases
APC
What is PTEN?
a phosphatase that dephosphorylates PIP3 back into PIP2