Paper

Rapid SARS-CoV-2 Adaptation to Available Cellular Proteases

Hoffmann, Markus, Kim, Yeonsu, Kröger, Andrea, Chaudhry, M Zeeshan, Abassi, Leila, Brunotte, Linda, Cicin-Sain, Luka, Grashoff, Martina, Ludwig, Stephan, Klawonn, Frank, Eschke, Kathrin, Pöhlmann, Stefan

2021

Overview

ABSTRACT Since the pandemic spread of SARS-CoV-2, the virus has exhibited remarkable genome stability, but recent emergence of novel variants show virus evolution potential. Here we show that SARS-CoV-2 rapidly adapts to Vero E6 cells that leads to loss of furin cleavage motif in spike protein. The adaptation is achieved by asymptotic expansion of minor virus subpopulations to dominant genotype, but wildtype sequence is maintained at low percentage in the virus swarm, and mediate reverse adaptation once the virus is passaged on human lung cells. The Vero E6-adapted virus show defected cell entry in human lung cells and the mutated spike variants cannot be processed by furin or TMPRSS2. However, the mutated S1/S2 site is cleaved by cathepsins with higher efficiency. Our data show that SARS-CoV-2 can rapidly adapt spike protein to available proteases and advocate for deep sequence surveillance to identify virus adaptation potential and novel variant emergence. Significance Statement Recently emerging SARS-CoV-2 variants B1.1.1.7 (UK), B.1.351 (South Africa) and B.1.1.248 (Brazil) harbor spike mutation and have been linked to increased virus pathogenesis. The emergence of these novel variants highlight coronavirus adaptation and evolution potential, despite the stable consensus genotype of clinical isolates. We show that subdominant variants maintained in the virus population enable the virus to rapidly adapt upon selection pressure. Although these adaptations lead to genotype change, the change is not absolute and genome with original genotype are maintained in virus swarm. Thus, our results imply that the relative stability of SARS-CoV-2 in numerous independent clinical isolates belies its potential for rapid adaptation to new conditions. Competing Interest Statement The authors have declared no competing interest. Footnotes * Classification Biological Sciences, Microbiology * Further experiments showed that although HS antagonists have an influence on the growth of low-passage SARS-CoV-2 strain on Vero E6 cells, these antagonist fail to inhibit S1/S2 site adaptation. Furthermore, new peptide cleavage data suggest that S1/S2 site adaptation in the Vero E6 cells is largely to optimize the cleavage by available proteases.

• Publisher: Cold Spring Harbor Laboratory Press,Cold Spring Harbor Laboratory
• Subject: Adaptation / Cathepsins / Clinical isolates / Coronaviruses / Furin / Genomes / Genotype & phenotype / Genotypes / Microbiology / Pandemics / Severe acute respiratory syndrome coronavirus 2 / Spike protein / Viruses