25-30 August 2019
Henry Ford Building
Europe/Berlin timezone

Multivalency and phase separation of measles virus replication machinery.

27 Aug 2019, 16:50
25m
Lecture Hall A (Henry Ford Building)

Lecture Hall A

Henry Ford Building

Talk Biological applications Biomolecules

Speaker

Serafima Guseva (Viral Replication Machines Group & Protein Dynamics and Flexibility by NMR Group, Institut de Biologie Structurale (IBS), CEA, CNRS, University Grenoble Alpes, Grenoble, France)

Description

RNA viruses tend to concentrate their replication machinery within so called “viral factories”. This membraneless compartment, formed by phase separation, has liquid properties and has been shown for several viruses in vivo [1,2]. Measles virus (MeV) is the cause of measles, it infects T-cells and macrophage cells, belongs to Paramyxoviridae family. Its genome consists of non-segmented negative-strand RNA, which encodes eight proteins from six genes (N, P, L, H, F and M). The MeV replication machinery consists of viral RNA covered by nucleoproteins (N), and protecting the genome against the host cell immune system, Large protein (L), the viral RNA-polymerase, and its essential co-factor, Phosphoprotein (P) [3]. As has been shown for other single strand RNA viruses there are N and P are essential for phase separation [1,2]. As shown by NMR, N and P interact with each other via long intrinsically disordered domains during their viral cycle [4,5].
Here we show for the first time phase separation of MeV N and P proteins in vitro, identify phase separation scaffold and characterise its liquid behaviour. Using NMR we describe multiple interactions between P and N disordered regions which are essential for phase transition and regulation of its biophysical parameters. In addition, we could show a non-stationary stoichiometry between N and P within droplets and propose the model on the basis of observations.
[1] Brunel J. et al., J Virol., 2014, 88(18):10851-63

[2] Nikolic J. et al., Nat Com, 2017, 8(58):00102-9

[3] Heinrich B. et al., mBio, 2018, 9(5):02290-17

[4] Jensen et al., PNAS, 2011, 108(24):9839-44

[5] Milles et al., Sci Adv, 2018, 22;4(8):eaat7778

Primary authors

Serafima Guseva (Viral Replication Machines Group & Protein Dynamics and Flexibility by NMR Group, Institut de Biologie Structurale (IBS), CEA, CNRS, University Grenoble Alpes, Grenoble, France) Dr Sigrid Milles (Groupe Flexibilité et Dynamique des Protéines par RMN, Institut de Biologie Structurale (IBS), CEA, CNRS, University Grenoble Alpes, Grenoble, France) Damien Maurin (Groupe Flexibilité et Dynamique des Protéines par RMN, Institut de Biologie Structurale (IBS), CEA, CNRS, University Grenoble Alpes, Grenoble, France) Dr Malene Ringkjobing Jensen (Groupe Flexibilité et Dynamique des Protéines par RMN, Institut de Biologie Structurale (IBS), CEA, CNRS, University Grenoble Alpes, Grenoble, France) Prof. Rob Ruigrok (Viral Replication Machines Group, Institut de Biologie Structurale (IBS), Univ. Grenoble Alpes, CEA, CNRS, Grenoble 38000, France) Dr Martin Blackledge (Groupe Flexibilité et Dynamique des Protéines par RMN, Institut de Biologie Structurale (IBS), CEA, CNRS, University Grenoble Alpes, Grenoble, France)

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