Sustained translational repression by eIF2α-P

mediates prion neurodegeneration

(Moreno, Mallucci et al.) (2012)

Luke Daly-Ronayne
Proteostasis
• Need to have the right amounts of functional protein in the cell at any
given time
• Maintaining amount requires balance of synthesis to degradation
• Maintaining functional protein requires correct initial folding during
synthesis and maintenance of conformation after synthesis
• Stress conditions impact both of these criteria (viral infection,
oxidative stress, amino acid starvation, and unfolded proteins)
• Under these conditions, translation is halted until the stress can be
addressed
Translation Initiation
• 2 main points of
intervention
• eIF4E phosphorylation
prevents recruitment to the
m7G cap
• eIF2α phosphorylation
prevents assembly of the
ternary complex (eIF2 +
initiator tRNA)
• eIF2α-P is how cells inhibit
translation under stress
conditions
(Figure source: Wheeler (2005); The process of initiation
of translation in eukaryotes)
The Unfolded Protein Response (UPR)

• Bip (chaperone) has two roles in UPR:

• (1) Binds to intracellular domains of UPR
kinase PERK to keep it in inactivated
• (2) Binds to unfolded proteins (exposed
hydrophobic residues, unpaired cysteines
etc.)
• Too many unfolded proteins = dissociation of
Bip from PERK
• PERK dimerises and autophosphorylates,
then phosphorylates eIF2α to eIF2α-P
The Problem with Prions
• Prions display strong misfolded features (side chains sticking out can
be packed with hydrophobic residues and unpaired cysteines)
• Once aggregation begins, actually breaking down the aggregates can
be very difficult
• This leads to chronic UPR activation, which diminishes global rates of
translation (bad for neuronal survival) and activates pro-apoptotic
pathways (also bad)
Decline of Synaptic Proteins
Reduces Synaptic Activity and
Eventually Kills Neurons
• Prion infection takes hold post-9wks
• Reduction in synaptic density,
presynaptic protein(?), postsynaptic
protein, excitatory postsynaptic
currents, burrowing behaviour
(quantifiable behavioural assay that’s
sensitive to prion disease), and number
of neurons in CA1 hippocampal region
• Also see spongiform pathology at wk9,
motor loss and death ~wk12
Prions Induce the UPR and
Repress Translation
• PrPSc is sensitive to Proteinase K (PK)
• PERK-P and eIF2α-P levels rise
• GADD34 doesn’t change, CHOP
expression spikes and caspase-12 is
cleaved
• Global translation drops (35S-Met
incorporation and polysome profiling)
• Increased ATF4 (response to attenuated
translation)
Treatment Scheme
Lowering eIF2α-P Levels Rescues
Neurons and Synaptic Function,
Increasing Them Does the
Opposite
• PrP(?), PERK-P and eIF2α-P levels
dropped by shPrP and GADD34,
increased by salubrinal
• Same pattern for other metrics (pre- and
post-synaptic density, EPSCs, burrowing,
synaptic density and neuronal count)
• Salubrinal omitted from some figures,
mentioned in-text to be worse than prion
only
Summary
• Prions induce the
UPR and repress
translation
• Targeting PrP itself
can be beneficial,
but so can a more
generalised
proteostastic
approach
(GADD34).
Implications for
therapeutic
development
Questions?