Principal Investigators
Dr. Maria Rosenthal & Prof. Dr. Charlotte Uetrecht
CSSB – Center for Structural Systems Biology
Bernhard Nocht Institute for Tropical Medicine &
Universität zu Lübeck
C8.2
PhD candidate
Morten Schumann
C8.2
Project Summary
Towards assembly and remodelling of Lassa virus ribonucleoparticles in vitro
Lassa virus is a hemorrhagic fever virus occasionally leading to outbreaks in west Africa against which no therapeutics or vaccines are available [1]. Lassa virus encodes only four structural proteins. The nucleoprotein NP and the polymerase L associate with the viral single-stranded RNA genome to form the ribonucleoprotein complex (RNP). The RNP serves as the functional unit of replication and transcription, which is why it constitutes a relevant target for antiviral drugs. During transcription or replication, the RNP complex has to undergo extensive remodelling in order for the polymerase to access the genome, while the nucleoprotein maintains its protective function [2, 3]. This mechanism is poorly understood.
To study the remodelling reaction, an RNP model will be assembled in vitro. Native mass spectrometry (nMS) will be used to detect formation of protein-protein and protein-RNA complexes. The second aim will be the establishment of a polymerase reaction in nMS-compatible conditions. Combining the first two aims, nMS will be used to observe the formation and remodelling of complexes during polymerase activity on the reconstituted RNP.
References:
- Hallam HJ, Hallam S, Rodriguez SE, Barrett ADT, Beasley DWC, Chua A, Ksiazek TG, Milligan GN, Sathiyamoorthy V, Reece LM. Baseline mapping of Lassa fever virology, epidemiology and vaccine research and development. NPJ Vaccines. 2018 Mar 20;3:11. doi: 10.1038/s41541-018-0049-5.
- Brunotte L, Kerber R, Shang W, Hauer F, Hass M, Gabriel M, Lelke M, Busch C, Stark H, Svergun DI, Betzel C, Perbandt M, Günther S. Structure of the Lassa virus nucleoprotein revealed by X-ray crystallography, small-angle X-ray scattering, and electron microscopy. J Biol Chem. 2011 Nov 4;286(44):38748-38756. doi: 10.1074/jbc.M111.278838.
- Sänger L, Williams HM, Yu D, Vogel D, Kosinski J, Rosenthal M, Uetrecht C. RNA to Rule Them All: Critical Steps in Lassa Virus Ribonucleoparticle Assembly and Recruitment. J Am Chem Soc. 2023 Dec 27;145(51):27958-27974. doi: 10.1021/jacs.3c07325.