Membrane Domains and Viral Assembly


Our team study the respective roles of virus capsid or matrix proteins and host cells lipids and lipid organisation in the assembly and budding of RNA envelopped viruses.


We adress 5 main questions:

  1. Lipid sorting during HIV-1 Gag self-assembly.
  2. Single Gag dynamics during HIV-1 Gag assembly in CD4+ T-Cells.
  3. Cortical actin dynamic and membrane curvature during virus assembly.
  4. Role of matrix M protein and cellular co-factors in influenza virus assembly.
  5. HIV-1 single viral particles assembly monitored by atomic force microscopy.

































1.Lipid sorting during HIV-1 Gag self-assembly.

We have previously shown that MLV assembly is induced by interaction of Gag matrix domain (MA) with PS-PIP2 lipids of the plasma membrane. (Hamard-Péron et al., 2010 ; Hamard-Péron & Muriaux, 2011). Instead of assembling in pre-existing plasma membrane domains, such as “rafts”, we proposed that the self assembly of HIV-1 Gag will induce the generation of acidic lipids enriched nanodomains at the plasma membrane of the host cells. (Kerviel et al., 2013, Yandapalli et al., 2014 and Mariani et al., 2014).

Using in silico coarse grained mecular dynamics, we proposed a new model of interaction of HIV-1 matrix domain (MA) of Gag with lipid membranes. In this new model, the sn-2 acyl chain of the PIP2 stays within the lipid membrane and the MA is surrounded by PIP2 and PS, as it is the case for MLV. (Charlier et al., 2014). This was experimentally confirmed later on by new NMR data. (Mercredi et al., 2016)

 

In vitro, using full length purified Gag and model lipid membrane with tuned compostions to mimic either « ratfs » containing plasma membrane or inner leaflet of plasma membrane, we showed that HIV-1 Gag self-assembly induced the formation of PIP2/Cholesterol enriched nanodomains. We also showed that sphingomyelin was excluded from these Gag self-assembly generated lipid nanodomains.  (Yandrapalli et al., 2016).   

Back to the top































2. Single Gag dynamics during HIV-1 Gag assembly in CD4+ T-Cells.

We are monitoring the time-course of HIV-1 assembly in living CD4 T cells, by observing and quantifying hundreds of thousands of individual Gag molecules motions (Mariani et al., 2016, see cover on the right).

Using dedicated motion analysis and big data algorithms, we can build maps of densities, diffusion, trapping energies and we observe their temporal changes in and out the budding sites (Floderer et al., 2018)













(Collaboration with teams of JB Sibarita (IINS, Bordeaux), M. Dahan (Institut Curie, Paris) and J.B. Masson (Institut Pasteur)).

Back to the top


















3. Cortical actin dynamic and membrane curvature during virus assembly.

We have previously shown that the CD81 tetraspanin played a role in HIV-1 assembly in CD4 T cells (Grigorov et al., 2009) and that the clathrin mediated endocytosis played a role in CD4 cell to cell HIV-1 transmission (Bosch et al., 2008). We recently shown that one of the main regulator of cortical actin network and membrane curvature played a role in HIV-1 assembly and release in CD4 T cells (Thomas et al., 2015).


From Mariani et al., 2014.

Back to the top




















































4. Role of matrix M protein and cellular co-factors in influenza virus assembly.

We are studying the role played by matrix proteins (M1 and M2) in the plasma membrane assembly of H1N1 influenza virus. M1 was shown to be necessary for viral assembly. We have recently shown that an arginine triplet (R76/77/78) of M1 was necessary for its plasma membrane localisation and for viral infectivity (Collaboration O.Moncorgé, IRIM and P. Roingeard, Tours University). Meanwhile, we also defined a minimal system for non infectious virus like particle (VLP) (Kerviel, Dash et al., 2016).




























5. HIV-1 single viral particles assembly monitored by atomic force microscopy.

under construction

Back to the top

(Click on the link to access publication or abstract)

2018

  1. Floderer C., Masson JB., Boiley E., Georgeault S., Merida P., El Beheiry M., Dahan M., Roingeard P., Sibarita JB., Favard C., Muriaux D. Live single molecule microscopy of HIV-1 assembly in host T cells reveals a spatio-temporal effect of the viral genome. BioRxiv, 267930.


2016

  1. Yandrapalli N, Lubart Q, Tanwar HS, Picart C, Mak J, Muriaux D, Favard C. Self assembly of HIV-1 Gag protein on lipid membranes generates PI(4,5)P2/Cholesterol nanoclusters. Sci Rep.,23 ;6:39332.
  2. Ahi YS, Zhang S, Thappeta Y, Denman A, Feizpour A, Gummuluru S, Reinhard B, Muriaux D, Fivash MJ, Rein A. Functional Interplay Between Murine Leukemia Virus Glycogag, Serinc5, and Surface Glycoprotein Governs Virus Entry, with Opposite Effects on Gammaretroviral and Ebolavirus Glycoproteins. mBio,7(6), pii : e01985-16
  3. Kerviel A, Dash S, Moncorgé O, Panthu B, Prchal J, Décimo D, Ohlmann T, Lina B, Favard C, Decroly E, Ottmann M, Roingeard P, Muriaux D. Involvement of an Arginine Triplet in M1 Matrix Protein Interaction with Membranes and in M1 Recruitment into Virus-Like Particles of the Influenza A(H1N1)pdm09 Virus. PLoS One.11(11):e0165421
  4. Balme S, Coulon PE, Lepoitevin M, Charlot B, Yandrapalli N, Favard C, Muriaux D, Bechelany M, Janot JM. Influence of Adsorption on Proteins and Amyloid Detection by Silicon Nitride Nanopore. Langmuir. 32(35):8916-25.
  5. Mariani-Floderer C, Sibarita JB, Favard C,Muriaux DM. Hunting Down HIV-1 Gag Proteins at the Plasma Membrane of Human T Lymphocytes. AIDS Res Hum Retroviruses. 32(7):658-9. (cover page)
  6. Martinez E, Allombert J, Cantet F, Lakhani A, Yandrapalli N, Neyret A, Norville IH, Favard C, Muriaux D, Bonazzi M. Coxiella burnetii effector CvpB modulates phosphoinositide metabolism for optimal vacuole development. Proc Natl Acad Sci U S A. ;113(23):E3260-9.
  7. Mercredi PY, Bucca N, Loeliger B, Gaines CR, Mehta M, Bhargava P, Tedbury PR, Charlier L, Floquet N, Muriaux D, Favard C, Sanders CR, Freed EO, Marchant J, Summers MF. Structural and Molecular Determinants of Membrane Binding by the HIV-1 Matrix Protein. J Mol Biol. ;428(8):1637-55.

2015

  1. Rodríguez-Mora S, Mateos E, Moran M, Martín MÁ, López JA, Calvo E, Terrón MC, Luque D, Muriaux D, Alcamí J, Coiras M, López-Huertas MR. Intracellular expression of Tat alters mitochondrial functions in T cells : a potential mechanism to understand mitochondrial damage during HIV-1 replication. Retrovirology,12,78-102
  2. A. Thomas, C. Mariani-Floderer, M.-R. Lopez-Huertas, N. Gros, E. Hamard-Péron, C. Favard, T. Ohlmann, J. Alcami, and D. Muriaux. Involvement of the Rac1-IRSp53-Wave2-Arp2/3 signaling pathway in HIV-1 Gag particle release in CD4 T cells. J. Virol., 89(16), 8162-81
  3. Bernaud J, Castelnovo M, Muriaux D, Faivre-Moskalenko C. Atomic force microscopy : a tool to analyze the viral cycle. Med Sci., 31(5), 522-8.
  4. S Adjili, A Favier*, G Fargier, A. Thomas, J Massin, K Monier, C Favard, C Vanbelle, S Bruneau, N Peyriéras, C Andraud, D Muriaux*, M-T Charreyre*. *Corresponding authors. Biocompatible Photoresistant Far-Red Emitting Fluorescent Polymer Probes with Near-Infrared Two-Photon Absorption for Living Cell and Zebrafish Embryo Imaging. Biomaterials, 46, 70-81.

2014

  1. Mariani C, Desdouits M, Favard C, Benaroch P, Muriaux D. Role of Gag and lipids during HIV-1 assembly in CD4(+) T cells and macrophages. Front Microbiol, 5, 312-317.
  2. Yandrapalli N, Muriaux D, Favard C. Lipid domains in HIV-1 assembly. Front Microbiol, 5, 220-225.
  3. J.M. Escoffre, M. Hubert, J. Teissié, M.P. Rols, C. Favard. Evidence for electro-induced membrane defects assessed by lateral mobility measurement of a GPi anchored protein Eur. Biophys. J., 43(6-7), 277-86
  4. S Adjili, A Favier, J Massin, Y Bretonnière, W Lacour, Y-C Lin, E Chatre, C Place, C Favard, D Muriaux, C Andraud, M-T Charreyre. Synthesis of Multifunctional Lipid-Polymer Conjugates : Application to the Elaboration of Bright Far-Red Fluorescent Lipid Probes. RSC Adv., 4 (30), 15569-78.
  5. L. Charlier, M. Louet, L. Chaloin, P. Fuchs, J. Martinez, D. Muriaux, C. Favard* & N. Floquet*. Coarse Grained Simulations of the HIV-1 Matrix Protein Anchoring : revisiting its assembly on membrane domains. *co-last authors. Biophys. J.,106(3):577-85.
  6. C. Faivre-Moskalenko, A. Thomas, K. Tartour, J. Bernaud, Y. Beck, M. Iazykov, J. Danial, M. Lourdin, D. Muriaux* & M. Castelnovo*. RNA control of HIV-1 particle size polydispersity. *co-last authors. PLOS one, 24 ;9(1):e83874.

2013

  1. M. Castelnovo, D. Muriaux & C. Faivre-Moskalenko. Entropic control of particle sizes during viral self-assembly. New J. Physics, 15:035028.
  2. Kerviel A, Thomas A, Chaloin L, Favard C, Muriaux D. Virus assembly and plasma membrane domains : Which came first ? Virus Res 171 : 332-340.
  3. Fargier G, Favard C, Parmeggiani A, Sahuquet A, Mérezègue F, Morel A, Denis M, Molinari N, Mangeat PH, Coopman PJ, Montcourrier P. Centrosomal targeting of Syk kinase is controlled by its catalytic activity and depends on microtubules and the dynein motor. FASEB J, 27(1):109–122.

2012

  1. Escoffre JM, Nikolova B, Mallet L, Henri J, Favard C, Golzio M, Teissie J, Tsoneva I, Rols MP. New insights in the gene electrotransfer process : evidence for the involvement of the plasmid DNA topology. Curr Gene Ther., 12(5):417-22.

2011

  1. Berger G, Durand S, Fargier G, Nguyen XN, Cordeil S, Bouaziz S, Muriaux D, Darlix JL, Cimarelli A. APOBEC3A is a specific inhibitor of the early phases of HIV-1 infection in myeloid cells. PLoS Pathog ;7(9):e1002221.
  2. Hamard-Peron E, Muriaux D. Retroviral matrix and lipids, the intimate interaction. Retrovirology ; 8:15-38.
  3. Favard C, Wenger J, Lenne PF, Rigneault H. FCS diffusion laws in two-phase lipid membranes : determination of domain mean size by experiments and Monte Carlo simulations. Biophys J ; 100(5):1242-51.
  4. Escoffre JM, Portet T, Favard C, Teissié J, Dean DS, Rols MP. Electromediated formation of DNA complexes with cell membranes and its consequences for gene delivery. Biochim Biophys Acta ; 1808(6):1538-43.

2010

  1. Muriaux D, Darlix JL. Properties and functions of the nucleocapsid protein in virus assembly. RNA Biol ; 7(6):744-53.
  2. Hamard-Peron E, Juillard F, Saad JS, Roy C, Roingeard P, Summers MF, Darlix JL, Picart C, Muriaux D. Targeting of murine leukemia virus gag to the plasma membrane is mediated by Pi(4,5)P2/P3 and a polybasic region in the matrix. J Virol ; 84(1):503-15.

2009

  1. Gasecka A, Han TJ, Favard C, Cho BR, Brasselet S. Quantitative imaging of molecular order in lipid membranes using two-photon fluorescence polarimetry. Biophys J ; 97(10):2854-62.
  2. Grigorov B, Attuil-Audenis V, Perugi F, Nedelec M, Watson S, Pique C, Darlix JL, Conjeaud H and D.Muriaux A role for CD81 on the late steps of HIV-1 replication in a chronically infected T cell line. Retrovirology ; 6:28.
  3. F.Perugi, D.Muriaux, B.C.Ramirez, S.Chabani, E.Decroly, JL.Darlix, V.Blot & C.Pique Human Discs Large is a new negative regulator of HIV-1 infectivity. Mol.Cell.Biol. ; 20:498-508.
  4. Taulet N, Comunale F, Favard C, Charrasse S, Bodin S, Gauthier-Rouvière C. N-cadherin/p120 catenin association at cell-cell contacts occurs in cholesterol-rich membrane domains and is required for RhoA activation and myogenesis. J Biol Chem ; 284(34):23137-45.
  5. Portet T, Camps i Febrer F, Escoffre JM, Favard C, Rols MP, Dean DS. Visualization of membrane loss during the shrinkage of giant vesicles under electropulsation. Biophys J ; 96(10):4109-21.

2008

  1. Macia E, Partisani M, Favard C, Mortier E, Zimmermann P, Carlier MF, Gounon P, Luton F, Franco M. The pleckstrin homology domain of the Arf6-specific exchange factor EFA6 localizes to the plasma membrane by interacting with phosphatidylinositol 4,5-bisphosphate and F-actin. J Biol Chem ; 283(28):19836-44.
  2. Fritz JV, Didier P, Clamme JP, Schaub E, Muriaux D, Cabanne C, Morellet N, Bouaziz S, Darlix JL, Mély Y, de Rocquigny H. Direct Vpr-Vpr interaction in cells monitored by two photon fluorescence correlation spectroscopy and fluorescence lifetime imaging Retrovirology ; 22, 87.
  3. Bosch, B., Grigorov, B., JL. Darlix, D. Muriaux & J. Esté. A clathrin-dynamin-dependent endocytic pathway for the uptake of HIV-1 by direct T cell-T cell transmission. Antiviral Research ; 80:185-93.
  4. L. Houzet, Z. Morichaud, L. Didier, D. Muriaux, JL. Darlix & M.Mougel. Deletion in nucleocapsid turn HIV-1 into a DNA-containing virus. Nucleic Acids Res ; 36, 2311-9.
  5. Corbin, B. Grigorov, P. Roingeard, J.-L. Darlix et D. Muriaux. Une nouvelle vision de l’assemblage du HIV-1 / Revisiting HIV-1 assembly. Médecines & Sciences ; 24:49-55.

Team Members




EnregistrerEnregistrerEnregistrerEnregistrerEnregistrerEnregistrerEnregistrerEnregistrer

Team Leader

Delphine Muriaux

Tenure Researcher CNRS, Virologist
More


EnregistrerEnregistrerEnregistrerEnregistrerEnregistrer

At a Glance

We aim at characterizing the respective roles of viral proteins and host-cell lipids or sub-plasma membrane constituents during the assembly of enveloped viruses affecting human beings (such as HIV-1 and influenza viruses).

We are biologists and biophysicists using multi-disciplinary approaches.

EnregistrerEnregistrer

Financial Supports

                                        
EnregistrerEnregistrerEnregistrerEnregistrerEnregistrer

Ecrivez-nous


     
       

   

Enregistrer

Enregistrer

Enregistrer

Enregistrer

Enregistrer

IRIM
Institut de Recherche en Infectiologie de Montpellier
UMR 9004 - CNRS / UM
1919 route de Mende - 34293 Montpellier cedex 5
FRANCE

 

Enregistrer

Enregistrer

Enregistrer

Enregistrer

Enregistrer

Enregistrer