| Abstract [eng] |
In the face of still ongoing COVID-19 diagnoses and mortality rates around the world, the development of drugs capable of targeting all SARS-CoV-2 strains is crucial, given the constant mutations and potential emergence of more dangerous virus variants. To address this, identifying a conserved protein target that remains unaffected by variation between different virus strains is essential. One such target is the helicase nsp13, a key enzyme involved in viral replication. By investigating the mechanical properties of SARS-CoV-2 helicase nsp13, our study aimed to deepen the understanding of its role within the replication complex. We successfully expressed and purified the nsp13 protein, qualitatively assessed its ATPase and helicase activity, and examined the impact of nsp13 on the polymerase nsp12 functionality. Notably, we observed that the interaction between purified nsp13 and the polymerase complex nsp12-nsp8-nsp7 did not significantly affect ATPase activity. However, this interaction significantly increased the polymerase activity of nsp12. Intriguingly, even when the active center amino acids of the nsp13 helicase (D374/E375) were replaced by alanines, the enhancement in polymerase activity was still observed. |
