Functional genomics screen identifies proteostasis targets that modulate prion protein (PrP) stability |
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| Authors: | Jennifer Abrams Taylor Arhar Sue Ann Mok Isabelle R Taylor Martin Kampmann Jason E Gestwicki |
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| Institution: | 1.Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158 USA ;2.Institute for Neurodegenerative Disease, University of California, San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158 USA ;3.Department of Biochemistry & Biophysics, University of California, San Francisco, San Francisco, CA 94158 USA |
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| Abstract: | Prion protein (PrP) adopts either a helical conformation (PrPC) or an alternative, beta sheet-rich, misfolded conformation (PrPSc). The PrPSc form has the ability to “infect” PrPC and force it into the misfolded state. Accumulation of PrPSc is associated with a number of lethal neurodegenerative disorders, including Creutzfeldt-Jacob disease (CJD). Knockout of PrPC protects cells and animals from PrPSc infection; thus, there is interest in identifying factors that regulate PrPC stability, with the therapeutic goal of reducing PrPC levels and limiting infection by PrPSc. Here, we assembled a short-hairpin RNA (shRNA) library composed of 25+ shRNA sequences for each of 133 protein homeostasis (aka proteostasis) factors, such as molecular chaperones and co-chaperones. This Proteostasis shRNA Library was used to identify regulators of PrPC stability in HEK293 Hu129M cells. Strikingly, the screen identified a number of Hsp70 family members and their co-chaperones as putative targets. Indeed, a chemical pan-inhibitor of Hsp70s reduced PrPC levels and limited conversion to PrPSc in N2a cells. These results implicate specific proteostasis sub-networks, especially the Hsp70 system, as potential new targets for the treatment of CJD. More broadly, the Proteostasis shRNA Library might be a useful tool for asking which proteostasis factors are important for a given protein.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12192-021-01191-8. |
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| Keywords: | shRNA Neurodegeneration Heat shock protein 70 Drug targets Chaperone networks Prion protein (PrP) |
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