Perrino, Giansimone
Wilson, Cathal
Santorelli, Marco
di Bernardo, Diego
2019-04-16
<p><strong>Highlights</strong></p>
<p>• <em>In silico</em> feedback control enables regulation of α-synuclein expression in yeast</p>
<p>• α-Synuclein inclusion formation is strictly concentration, but not time, dependent</p>
<p>• The aggregation threshold of the α-synuclein A53T mutant is 56% of the wild-type</p>
<p>• Autophagy induction speeds up inclusion clearance in the A53T α-synuclein strain</p>
<p><strong>Summary</strong></p>
<p>Aggregation of α-synuclein and formation of inclusions are hallmarks of Parkinson’s disease (PD). Aggregate formation is affected by cellular environment, but it has been studied almost exclusively in cell-free systems. We quantitatively analyzed α-synuclein inclusion formation and clearance in a yeast cell model of PD expressing either wild-type (WT) α-synuclein or the disease-associated A53T mutant from the galactose (Gal)-inducible promoter. A computer-controlled microfluidics device regulated α-synuclein in cells by means of closed-loop feedback control. We demonstrated that inclusion formation is strictly concentration dependent and that the aggregation threshold of the A53T mutant is about half of the WT α-synuclein (56%). We chemically modulated the proteasomal and autophagic pathways and demonstrated that autophagy is the main determinant of A53T α-synuclein inclusions’ clearance. In addition to proposing a technology to overcome current limitations in dynamically regulating protein expression levels, our results contribute to the biology of PD and have relevance for therapeutic applications.</p>
https://doi.org/10.1016/j.celrep.2019.03.081
oai:zenodo.org:2668451
Zenodo
https://zenodo.org/communities/cosy-bio
https://zenodo.org/communities/eu
info:eu-repo/semantics/openAccess
Creative Commons Attribution 4.0 International
https://creativecommons.org/licenses/by/4.0/legalcode
Cell Reports, 27(3), 916-927, (2019-04-16)
Bioengineering
Microfluidics
Feedback control
Gene expression
Synuclein
Aggregation
Quantitative Characterization of α-Synuclein Aggregation in Living Cells through Automated Microfluidics Feedback Control
info:eu-repo/semantics/article