Open AccessFunctional genomics screen identifies proteostasis targets that modulate prion protein (PrP) stability
Author(s) -
Jennifer L. Abrams,
Taylor Arhar,
Sue-Ann Mok,
Isabelle R. Taylor,
Martin Kampmann,
Jason E. Gestwicki
Publication year - 2021
Publication title -
cell stress and chaperones
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.994
H-Index - 87
eISSN - 1466-1268
pISSN - 1355-8145
DOI - 10.1007/s12192-021-01191-8
Subject(s) - proteostasis , biology , small hairpin rna , microbiology and biotechnology , neurodegeneration , protein folding , rna , genetics , gene , disease , medicine , pathology
Prion protein (PrP) adopts either a helical conformation (PrP C ) or an alternative, beta sheet-rich, misfolded conformation (PrP Sc ). The PrP Sc form has the ability to "infect" PrP C and force it into the misfolded state. Accumulation of PrP Sc is associated with a number of lethal neurodegenerative disorders, including Creutzfeldt-Jacob disease (CJD). Knockout of PrP C protects cells and animals from PrP Sc infection; thus, there is interest in identifying factors that regulate PrP C stability, with the therapeutic goal of reducing PrP C levels and limiting infection by PrP Sc . 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 PrP C 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 PrP C levels and limited conversion to PrP Sc 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.