A yeast-based system to study SARS-CoV-2 M pro structure and to identify nirmatrelvir resistant mutations
Year: 2022
Type: preprint
Abstract: The SARS-CoV-2 main protease (M pro ) is a major therapeutic target. The M pro inhibitor, nirmatrelvir, is the antiviral component of Paxlovid, an orally available treatment for COVID-19. As M pro inhibitor use increases, drug resistant mutations will likely emerge. We have established a non-pathogenic system, in which yeast growth serves as a proxy for M pro activity, enabling rapid identification of mutants with altered enzymatic activity and drug sensitivity. The E166 residue is known to be a potential hot spot for drug resistance and yeast assays showed that an E166R substitution conferred strong nirmatrelvir resistance while an E166N mutation compromised activity. On the other hand, N142A and P132H mutations caused little to no change in drug response and activity. Standard enzymatic assays confirmed the yeast results. In turn, we solved the structures of M pro E166R, and M pro E166N, providing insights into how arginine may drive drug resistance while asparagine leads to reduced activity. The work presented here will help characterize novel resistant variants of M pro that may arise as M pro antivirals become more widely used.
Language: English
Subfield: Infectious Diseases
Field: Medicine
Domain: Health Sciences
Open Access status: green
Awards: G1678787968, G1756342139, Phase II SER-CAT Optimization: Acquisition of a next generation area detector, G2123841078, G3338542329, Acquisition of a High Precision Micro Diffractometer to Support NIH funded Research at SER-CAT, G434509735, G572720644, G5921955326, Heterogeneous synthetic microbiome constructed with biopolymer fluitrodes, G760289873, G7932540327, G9000285124, Development of dual inhibitors targeting the viral main protease and the host cathepsin L as SARS-CoV-2 antivirals, Acquition of a microdiffractometer for SER-CAT
OpenAlex ID: W4290988437