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Honours Project in the Peet Lab 2014
O2-dependent regulation of TRP Ion Channels
Our research focus is to understand how cells sense and respond to changes in oxygen.
Oxygen sensing, and more importantly the response to low oxygen or hypoxia, is essential
for survival, with direct relevance to normal physiology and development. The response to
hypoxia is also an essential part of most major human diseases, including ischaemic (eg.
heart attack, stroke, etc.), vascular, and metabolic diseases, and cancer. Not surprisingly,
this is a hot topic of international research.
Many of the responses to hypoxia involve changes in gene expression, and these
responses are mediated by a novel class of oxygen-sensing enzymes. These enzymes
are hydroxylases, one of which is the asparaginyl hydroxylase Factor Inhibiting HIF (FIH).
They catalyse the oxygen-dependent post-translation modification of the HIF transcription
factors which alter their activity and in turn regulate gene expression. We and others have
shown that FIH plays an important role in metabolism, and has been implicated in cancer.
We have also shown that FIH can modify other proteins and pathways, although the
function and consequence of these modifications so far remains unclear.
We have recently identified the TRP ion channels as substrates of enzyme FIH. These
cation channels have a number of different functions, including sensing temperature and
pain, and are linked to a range of human diseases. Our preliminary data demonstrate that
at least one of these channels, TRPV3, is hydroxylated by FIH. Importantly, we have also
demonstrated that this modification modifies channel activity (current flow) in an oxygen
dependent manner.
O2
FIH
H
-OH
?
This exciting Honours project will characterise how hydroxylation alters channel activity,
whether other closely related channels can also be regulated by FIH, and how this relates
to normal channel function and disease. The project will involve a range of molecular and
biochemical techniques, including subcloning, PCR, protein expression, western blotting,
and mammalian cell culture. This is a collaborative project with Dr Grigori Rychkov
(expertise in ion channels, patch clamping) from the Discipline of Physiology and SAHMRI,
providing an opportunity to gain experience in related techniques, such as patch clamping
and the analysis of channel function in animal models.
If you have any questions regarding this project, I would be more than happy to discuss
them with you.
Dan
Dan Peet
Room 3.40 MLS Building
Phone: 8313 5367
Email: dan.peet@adelaide.edu.au
Web site: https://www.adelaide.edu.au/mbs/research/peet/