RTK structure - structure and function of the non-structural proteins in SARS-CoV-2 replication and transcription complexes (RTK-Struktur)

The overall goal of the project is to transfer our work on SARS-CoV non-structural proteins (NSP) to SARS-CoV-2. For this purpose, all NSPs that are not membrane-bound should be expressed both in bacteria and in eukaryotic systems. Subsequently, partial complexes of the replication and transcription complexes (RTK) should be assembled from the purified NSP and checked for their functionality, e.g. RNA binding and processing. After characterization with native mass spectrometry (MS), selected complexes are to be fed into functional assays and structural biological methods with higher resolution in order to obtain a better understanding of virus biology. This application relates to the announcement "Funding call for research into COVID-19 in the course of the Sars-CoV-2 outbreak of 03.03.2020". According to the announcement, the subject of funding is to investigate the biology of the virus. The work is carried out as part of Module 4 - Support for ongoing research projects on coronaviruses - and extends the ERC Starting Grant ERC-StG-2017 759661 "SPOCk'S MS - Sampling Protein cOmplex Conformational Space with native top down Mass Spectrometry". The goals are as follows: - Expression of the SARS-CoV-2 NSPs - Assembly of protein complexes of the RTKs - Structural and functional study of the complexes obtained