Laboratory name: Laboratoire de Physique, ENS de Lyon
CNRS identification code: UMR 5672
Supervisors : Fabien Montel and Cendrine Moskalenko
e-mail:
Fabien.montel@ens-lyon.fr,
cendrine.moskalenko@ens-lyon.fr Phone number: +33 (0)4 26 23 38 23
Web page: [
perso.ens-lyon.fr]
Interaction and transport of viruses through the nuclear pore
Each virus family has its own mode of disassembly and release of the viral genome. For some viruses, this step can only take place after the viral capsid has entered the cell nucleus through the nuclear pore [1], a biological nanopore that regulates exchanges between the nucleus and the cytoplasm. We have also shown that this pore has plasticity to adapt to the transcriptional state of the cell [2].The general objective of this collaborative project between two teams at the physics-biology interface is to characterize the physical (geometry, mechanics) and biological (specific peptide motif) factors that control this transport. In particular, we will characterise the energetic landscape of virus entry into the nucleus.
This project will focus on two viruses that transit the nuclear pore and that we are currently studying in the team [3]: a gene therapy vector (Adeno Associated Virus, AAV) and the Hepatitis B virus (HBV). In this project we will implement a physical characterization of nuclear translocation processes in controlled systems, in order to study and model the impact of nano-mechanical properties of viral particles in this phenomenon. To do this, we will use Atomic Force Microscopy (AFM). It allows us to probe the morphology of biological objects (capsids, nuclear pore) at a very high spatial resolution and also to exert mechanical constraints on these objects in order to measure their elastic response [4]. We will also use nanofluidic tools to measure the transport of viral particles through the nuclear pore.
First, in a simplified and controlled system, we will measure the topography and nano-mechanics of nuclear envelopes of Xenope, a membrane containing many nuclear pores, by AFM in liquid medium [5]. After analyzing the mechanical contribution of each part of the nuclear pore, the envelopes will be placed in the presence of viruses (AAV and HBV). The location of the viruses in relation to the nuclear pore and their contribution to the nano-mechanics of the pore will then be determined.
In a second step, we will measure the transport properties of the viruses through these nuclear envelopes using an optical near-field system developed in the team (zero mode wave guide for nanopores [6]). This methodology will allow us to quantify the translocation energy landscape at the single virus scale in real time.
References :
[1] S.R. Wente & M.P. Rout, Cold Spring Harb Perspect Biol 2010.
[2] J. Sellés, M. Penrad-Mobayed, C. Guillaume, A. Fuger, L. Auvray, O. Faklaris & F. Montel, Scientific Reports 2017, 14732
[3] J. Bernaud, A Rossi, A Fis, L Gardette, L Aillot, H Büning, M Castelnovo A. Salvetti & C. Faivre-Moskalenko, J. Biol. Phys. 2018, 44 (2), 181-194
[4] W.H. Roos, R. Bruinsma & G.J.L. Wuite; Nature Physics 2010, 6, 633.
[5] G.J. Stanley, A. Fassati & B.W. Hoogenboom, Life Sci. Alliance 1 2018, e201800142
[6] T. Auger, J. Mathé, V. Viasnoff, G. Charron, J.-M. Di Meglio, L. Auvray & F. Montel, Phys Rev Lett. 2014, 113, 028302.