Freezing dynamics of SARS-CoV-2 replication-transcription machinery using time-resolved cryo-EM

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused tremendous effects on global health the last two years, and there is a high risk of new coronavirus outbreaks in the future.

The replication-transcription complex (RTC) of coronaviruses is a highly dynamic and multifunctional machinery and essential for virus proliferation.

The RTC undergoes a number of conformational changes during RNA synthesis but structural information is currently limited to snapshots of a few distinct conformations that the protein complex explores at equilibrium, and the exact details of the molecular mechanisms remain elusive.

Thus, I propose to use time-resolved cryo-electron microscopy to capture the short-lived active intermediate conformations of the SARS-CoV-2 RNA synthesis.

To realise the proposed project, I will spend two years at the Deutsches-Elektronen Synchrotron (DESY) and the labs of Professors Henry Chapman and Charlotte Uetrecht. For my final year, I am moving to Uppsala University and the lab of Professor Filipe Maia. The main goal of the proposed research is to advance understanding of how the RTC machinery is functioning.

Specifically, I will focus on the polymerase and exoribonuclease activity since current antiviral drugs that target the polymerase are less effective on coronaviruses due to the exoribonuclease. Better knowledge of the RTC may one day lead to design more effective antiviral drugs.