Nathalie SAUVONNET

CURRICULUM VITAE

NAME, Surname:SAUVONNET Nathalie

Professional address:Institut Pasteur, Unité de Pathogénie Microbienne Moléculaire

28 rue du Dr Roux Paris Cedex 15

E-mail:

Position title:PhD, HDR, Directeur de Recherche, Group LeaderInstitut Pasteur

EDUCATION/TRAINING

1988:Baccalaureate Scientific, série C (Mathematics, Physics, Biology), Paris, France

1992:Bachelor of Biology and Applied Genetics (BGA), University Paris 7, France

1993:Master of BGA, University Paris 7, France. Training at University Paris-Sud, Orsay, Institute of Genetic and Microbiology,Pr. I. B. Holland. Title: «Study of Type I secretion of haemolysin in Escherichia coli»

1994:Master of Science in Microbiology, University Paris 7, France. Training at Institut Pasteur (IP), Paris, France, Molecular Genetic Unit, Pr. A. P. Pugsley. Title: “Type II secretion of pullulanase from Klebsiella oxytoca”

1998:PhD in Microbiology, University Paris 7 and IP, Paris, France, Molecular Genetic Unit, Pr. A. P. Pugsley. Title: “Type II secretion of pullulanase from Klebsiella oxytoca”

2009:HDR (Habilitation à Diriger la Recherche), University Paris 5.

POSITIONS AND HONORS

1994-1998:PhD student in Pr A. Pugsley’s lab at Institut Pasteur(IP), Paris. Awarded Postgraduate Scholarship of the Ministry of Research and Technics of France (MRT)

1998-2000:Postdoctorate in Pr A. Pugsley’s lab at IP, Paris. Awarded postdoctoral fellowship “Bourse Roux” from IP.“Type IV pili in Escherichia coli”

2000-2002:Postdoctorate in Pr G. Cornelis’s lab at Christian de Duve Institute of Cellular Pathology, Belgium. Awarded postdoctoral fellowship“Marie Curie” from the European Union (EU). "Role of Type III secreted factors of Yersinia"

2002-2003:Postdoctorate in Pr A. Dautry-Varsat’s lab(IP). Awarded postdoctoral fellowship from the ARC. “Endocytosis of cytokine receptors”

2003:Recruited as “Chargée de recherche” at IP

2005-today:Team leader of the "Intracellular traffic" team

2006-today:Member of “Comité d’Animation Scientifique (CAS)”, Committee organizing seminar and congresses for the department of Cellular Biology and Infection at IP.

2010:Member of the expertcommittee for the chairInserm-Université Paris 5-Descartes

2010-today:Board memberof the ClubExocytosis-Endocytosis and organizing meetings. Congress's organizer in 2009, 2011 and 2014.

2015:Group leader, Institut Pasteur

2015:Member of the "Bureau" of COMité d'Evaluation des activités Scientifiques des Personnels IP (COMESP)

2017:Elected Member of "Assemblée des 100" of Institut Pasteur

2017:Member of PPU committee

TEACHING

2005: Master of Ecole Normale Supérieur (ENS) Paris. 3h courses «Endocytosis and intracellular trafficking»

2006:Master of Molecular Biology of the Cell at University Paris 6. 3h courses«Endocytosis and intracellular trafficking»

2006-today: Master of Physiology, Cytoskeleton and Intracellular Traffickingat University Paris 5. Each year 3h courses «Endocytosis and intracellular trafficking».

2008: EMBO Practical Courses «Cell biology of host-pathogens interactions» at Institut Pasteur on «Endocytosis by using Total Internal Fluorescence Microscopy».

2009: Pasteur-Asia Cell Biology Course at Hong Kong university in China. 3h courses «Endocytosis and intracellular trafficking»

2010-2011-2015: Practical Courses in Master of Molecular Biology of the Cell at Institut Pasteur- Institut Curie. Each year, one week training on «Imaging and analyzing receptor-mediated endocytosis»

2011:Workshop on Image Analysis, from Theory to Application at Institut Pasteur on «Colocalization: towards a reliable method?».

EXPERTISE AND ASSESSMENTS

External reviewer or "rapporteur" for doctoral /HDR dissertation:

2010: External reviewer for thePhD Maria Dolores Fernandez Garcia (University Paris 7)

2011: PhD Naga Sailaja Imjeti (University Paris 11).

2012: PhD Maddalena Costanzo (University Paris 11).

2014: HDR Stéphanie Lebreton (University Paris 11)

2016: PhD Florence Marie-Anais ,and Jean-Marie-Carpentier (University Paris Descartes)

2017: HDR Erwan Mortier (University of Nantes)

External reviewer for Master:

2011, 2015: Member of Jury of Master Molecular Biology of the Cell (IP/Institut Curie)

2012, 2013, 2014: Member of Jury of Master of Genetics (University Paris 7)

Referee for peer-reviewed journals: about 10 papers per year for several journals J Cell Sci, Traffic, Cellular Microbiol, Plos One, Science, MBoc

Speaker in several congresses and invited seminars: At the EMBO conference serieson Endocytosis in Greece and in Switzerland. At the International Jacques Monod Conference "Molecular basis for membrane remodeling and organization" in France and at the International Symposium in Switzerland. At the SBCF meeting “Building the cell” in Paris. Invited at the Nanoscience center, university of Jyvaskyla (Finland).

RESEARCH SUPERVISION

Director of Postdoctorate fellows:

2012-today: Mariana Ferrari. Project title: «Maintenance of organelle identity: Shigella as tool to decipher basic mechanisms of secretion in eukaryotic cells”. FRM fellowship.

2014-today: Alexandre Grassart. Project title: “Dynamics of the cytokine receptorinternalization in its native physiological environment.” Bourse Roux IP.

Director of PhD students:

2006-2010: Alexandre Grassart. Project title: «Clathrin-independent endocytosis: link betweenactin cytoskeleton andthe internalizationof interleukin 2 receptor”. Fellowship from the Ministry of Research and Technics of France (MRT), ED GGC.

2009-2013: Cyril Basquin. “Identification of specific factors involved in clathrin-independent endocytosis of cytokine receptors”. Fellowship from the Ministry of Research and Technics of France (MRT), ED GGC.

2013-2016: Laetitia Bertot. "Role of membrane bending proteins in clathrin-independent endocytosis of cytokine receptors". Fellowship from the Ministry of Research and Technics of France (MRT), ED GGC.

Director of Master students:

2011: Marie-Claire Mateo Master 1 of Cellular Biology University Paris 6. Project title:«Comparisonof endocytosisof Iota, the toxinof Clostridium,with the clathrin-independentuptake of Interleukin2 receptor”

2012:Lea Ripoll. Master 1 of Cellular Biology, Physiology and Pathology. University Paris 7. “Role of the WAVE complex in Interleukin2 receptor endocytosis”

2012-2013: Laetitia Bertot. Master 2 of Cellular Biology, University Paris 6. “Factors involved in membrane bending during clathrin-independent endocytosis”

RESEARCH SUPPORT

2010-2012: Awarded Pasteur-Weizmann Joint Research Program that funds two groups at IP and Weizmann (IW)Principal investigator: Nathalie Sauvonnet (IP) and Nir Gov (IW): “Clathrin-independent endocytosis of receptors driven by the actin cytoskeleton”

2011-2014: Awarded Programme Transversal de Recherche (PTR387)Principal investigator: Nathalie Sauvonnet (IP) Collaborators: Jean-Christophe Olivo-Marin’s group (IP) and Philippe Chavrier’s group (Institut Curie): “Computational imaging of clathrin-dependent and -independent endocytosis dynamics: a comparative study of the roles of actin”

2012-2013: Awarded La ligue Contre le Cancer funding for the fourth year PhD of Cyril Basquin

2015-2018: LABEX IBEID with F Arenzana lab coordinator B Lagane.«Heterogeneity of CC Chemokine Receptor 5 (CCR5) conformations: Characterization in living cells, molecular mechanisms and consequences on HIV pathogenesis and inhibition»

2016-2018: Awarded Programme Transversal de Recherche (PTR) PI: Nathalie Sauvonnet " Reconstitution of Mycobacterium and Shigella infection in physiological condition using organ-in-a-chip "

2017-2020: DIM ELICIT PI: Samy Gobaa "Novel Organ-on-Chip Microfluidic device based on Hydraulically Actuated Hydrogel Layers"

PUBLICATIONS

  1. Lagache T, Grassart A, Dallongeville S, Faklaris O, Sauvonnet N, Dufour A, Danglot L, Olivo-Marin JC. 2018. Mapping molecular assemblies with fluorescence microscopy and object-based spatial statistics.Nat Commun. 2018 Feb 15;9(1):698
  1. Bertot L, Grassart A, Lagache T, Nardi G, Basquin C, Olivo-Marin JC, Sauvonnet N.2018.Quantitative and Statistical Study of the Dynamics of Clathrin-Dependent and -Independent Endocytosis Reveal a Differential Role of EndophilinA2.Cell Rep. 2018 Feb 6;22(6):1574-1588
  1. Bonvicini A, Guilhaudis L, Tognetti V, Desmaële D, Sauvonnet N, Oulyadi H, Joubert L. 2018. Revisiting absorption and electronic circular dichroism spectra of cholesterol in solution: a joint experimental and theoretical study.Phys Chem Chem Phys. 2018 Feb 14;20(7):5274-5284.
  1. Fernandez-Garcia MD, Meertens L, Chazal M, Hafirassou ML, Dejarnac O, Zamborlini A, Despres P, Sauvonnet N, Arenzana-Seisdedos F, Jouvenet N, Amara A. Vaccine and Wild-Type Strains of Yellow Fever Virus Engage Distinct Entry Mechanisms and Differentially Stimulate Antiviral Immune Responses. MBio. 2016 Feb 9;7(1):e01956-15.
  1. Basquin C, Trichet M, Vihinen H, Malardé V, Lagache T, Ripoll L, Jokitalo E, Olivo-Marin JC, Gautreau A, Sauvonnet N. 2015. Membrane protrusion powers clathrin-independent endocytosis of interleukin-2 receptor.EMBO J. 2015 Aug 13;34(16):2147-61
  1. Lagache T, SauvonnetN, Danglot L, Olivo-Marin JC. 2015. Statistical analysis of molecule colocalization in bioimaging.Cytometry A. 2015 Jan 20.
  1. Boucrot E, FerreiraA, Almeida-Souza A, Vallis Y, Howard G, DebardS, Bertot L, Sauvonnet N and McMahon HT. 2015"Endophilin marks and controls a clathrin-independent endocytic pathway" Nature. 2015 Jan 22;517(7535):460-5
  1. Boissan M, Montagnac G, Guitton J, Romao M, Sauvonnet N, Lagache T, Lascu I, Raposo G,Lacombe ML, Polo S, Roux Aand Chavrier P. A 2014 Nucleoside Diphosphate Kinase fuels Dynamin with GTP for efficient endocytosis. SCIENCE 2014 Jun 27;344(6191):1510-5
  1. Jin J, Colin P, Staropoli I, Lima-Fernandes E, Ferret C, Demir A, Rogee S, Hartley O, Randriamampita C, Scott M, Marullo S, Sauvonnet N, Arenzana-Seisdedos F, Lagane B and Brelot A. 2014Targeting Spare CCR5 as a Principle to Inhibit HIV-1 J Biol Chem. 2014 Jul 4;289(27):19042-52
  1. Lagache T, Lang G, Sauvonnet N and Olivo-Marin JC. 2013 Analysis of the Spatial Organization of Molecules with Robust StatisticsPLOS ONE, 2013 Dec 4;8(12)
  1. Basquin C, SauvonnetN. 2013. Phosphoinositide 3-kinase at the crossroad between endocytosis and signaling of cytokine receptors.Commun Integr Biol. Jul 1;6(4)

12.Basquin C, Malardé V, Mellor P,Anderson DA, Meas-Yedid V, Olivo-Marin JC, Dautry-Varsat, A, Sauvonnet N. 2013. The signaling factor PI 3-kinase is a specific regulator of the clathrin-independent dynamin-dependent endocytosis of IL-2 receptors. J Cell Sci. (126):1099-1108

  1. Wigelsworth DJ, Ruthel G, Schnell L, Herrlich P, Blonder J, Veenstra TD, Carman RJ, Wilkins TD, Van Nhieu GT, Pauillac S, Gibert M, SauvonnetN, Stiles BG, Popoff MR, Barth H. 2012.CD44 Promotes Intoxication by the Clostridial Iota-Family Toxins. PLoS One.;7(12):e51356.
  1. Mounier J, Boncompain G, Senerovic L, Lagache T, Chrétien F, Perez F, Kolbe M, Olivo-Marin JC, Sansonetti, P, Sauvonnet, N. 2012. Cholesterol relocation induced by the Shigella virulence factor IpaB inhibits host cell secretion by disrupting the Golgi complex and recycling network. Cell Host and Microbe, Sep 13;12(3):381-9.
  1. Grassart A, Meas-Yedid V, Dufour A, Olivo-Marin JC, Dautry-Varsat A, Sauvonnet N. 2010. Pak1 phosphorylation enhances cortactin-N-WASP interaction in clathrin-caveolin-independent endocytosis. Traffic. Aug;11(8):1079-91
  1. Meas-Yedid, V, Dufour, A, Zhang, B,Grassart, A, Sauvonnet N, and Olivo-Marin JC. 2010. Automated quantification cellular processes with applications to high-content screening. Handbook of Pattern Recognition & Computer (4 Edn). Edited by Prof. C.H. Chen, to be published by World Scientific Publishing January 2010 p. 537-547.
  1. Geny B, Grassart A, Manich M, Chicanne G, Payrastre B, Sauvonnet N, MR. Popoff. 2010. Rac1 inactivation by lethal toxin from Clostridium sordellii modifies Focal Adhesions upstream of actin depolymerization. Cell Microbiol. Feb;12(2):217-32.
  1. Antonescu CN, Foti M, Sauvonnet N and A Klip.(2009). Ready, Set, Internalize: Mechanisms and Regulation of GLUT4 Endocytosis. Bioscience Reports. Feb;29(1):1-11
  1. Grassart A, Dujeancourt A, Lazarow PB, Dautry-Varsat A, Sauvonnet N. 2008. Clathrin-independent endocytosis used by the IL-2 receptor is regulated by Rac1, Pak1 and Pak2. EMBO Rep. Apr;9(4):356-62.
  1. Sauvonnet N, Dujeancourt A, Dautry-Varsat A. 2005.Cortactin and dynamin are required for the clathrin-independent endocytosis of gammac cytokine receptor. J Cell Biol. Jan 3;168(1):155-63.
  1. Gesbert F, Sauvonnet N, Dautry-Varsat A. 2004. Clathrin-lndependent endocytosis and signaling of interleukin 2 receptors IL-2R endocytosis and signaling. Curr Top Microbiol Immunol. 286:119-48. Review.
  1. Sauvonnet N, Lambermont I, van der Bruggen P, Cornelis GR. 2002. YopH prevents monocyte chemoattractant protein 1 expression in macrophages and T-cell proliferation through inactivation of the phosphatidylinositol 3-kinase pathway. Mol Microbiol. Aug;45(3):805-15.
  1. Sauvonnet N, Pradet-Balade B, Garcia-Sanz JA, Cornelis GR. 2002. Regulation of mRNA expression in macrophages after Yersinia enterocolitica infection. Role of different Yop effectors. J Biol Chem. Jul 12;277(28):25133-42.
  1. Pugsley AP, Bayan N, Sauvonnet N. 2001. Disulfide bond formation in secreton component PulK provides a possible explanation for the role of DsbA in pullulanase secretion. J Bacteriol. Feb;183(4):1312-9.
  1. Sauvonnet N, Vignon G, Pugsley AP, Gounon P. 2000. Pilus formation and protein secretion by the same machinery in Escherichia coli. EMBO J. May 15;19(10):2221-8.
  1. Sauvonnet N, Gounon P, Pugsley AP. 2000. PpdD type IV pilin of Escherichia coli K-12 can Be assembled into pili in Pseudomonas aeruginosa. J Bacteriol. Feb;182(3):848-54.
  1. Guilvout I, Hardie KR, Sauvonnet N, Pugsley AP. 1999. Genetic dissection of the outer membrane secretin PulD: are there distinct domains for multimerization and secretion specificity? J Bacteriol. Dec;181(23):7212-20.
  1. Sauvonnet N, Pugsley AP. 1998. The requirement for DsbA in pullulanase secretion is independent of disulphide bond formation in the enzyme. Mol Microbiol. Feb;27(3):661-7.
  1. Pugsley AP, Francetic O, Possot OM, Sauvonnet N, Hardie KR. 1997. Recent progress and future directions in studies of the main terminal branch of the general secretory pathway in Gram-negative bacteria--a review. Gene. Jun 11;192(1):13-9. Review.
  1. Pugsley AP, Francetic O, Hardie K, Possot OM, Sauvonnet N, Seydel A. 1997. Pullulanase: model protein substrate for the general secretory pathway of gram-negative bacteria. Folia Microbiol (Praha).;42(3):184-92. Review.
  1. Sauvonnet N, Pugsley AP. 1996. Identification of two regions of Klebsiella oxytoca pullulanase that together are capable of promoting beta-lactamase secretion by the general secretory pathway. Mol Microbiol. Oct;22(1):1-7.
  1. Chervaux C, Sauvonnet N, Le Clainche A, Kenny B, Hung AL, Broome-Smith JK, Holland IB. 1995. Secretion of active beta-lactamase to the medium mediated by the Escherichia coli haemolysin transport pathway. Mol Gen Genet. Nov 15;249(2):237-45.
  1. Sauvonnet N, Poquet I, Pugsley AP. 1995 Extracellular secretion of pullulanase is unaffected by minor sequence changes but is usually prevented by adding reporter proteins to its N- or C-terminal end. J Bacteriol. Sep;177(18):5238-46.

ACHIEVEMENTS

Nathalie Sauvonnet research work has always been focused onprotein transportacross membranesleading totheirselectivesorting to theircompartments. These mechanisms ofintracellular proteintraffickingareessential for prokaryotesas well as for eukaryotes and define cellular organization. First, she studied these mechanismsin bacteriaand then in mammals.

In Gram-negative bacteria (K. oxytoca, E. coli, Y. enterocolitica), Nathalie Sauvonnet worked on secretion mechanisms (Type 1, 2, 3 secretion system, TnSS) and on the surface appendage, type IV pili. These pathways can be implicated in bacterial pathogenesis. She answered to several key issues:

- She showed that Pullulanase using T2SS, was one of the first examples of a folded protein during its secretion (21, 25, 30). This concept of folded protein during secretion has been generalized for T2SS.

- She identified a bipartite secretion signal of the T2SS substrate Pullulanase (28, 30). Since then, other works converge to the idea of a conformational signal gathering several motifs spread along the polypeptide sequence of the protein using T2SS.

- Strikingly, she showed that the T2SS machinery isassembledinpili-like bundles (21, 22). These results have openednew avenues ofinvestigations onthe role ofT2SS pili in the secretion, adhesion orpathogenicity. The T2SS pilus is now proposed to be a piston, pushing the secreted proteins through the outer membrane.

- She performed a transcriptomic analysis of host cell infected by several strains of Y. entrocolitica and showed that the main action of T3SS is to counteract the host cell pro-inflammatory response to infection (19, 20). Since then, this action of Yersinia has been also found in many other pathogens.

Cell compartmentalization in eukaryotes is very complex and tightly controlled. In Mammals, Nathalie Sauvonnet’s team is focused on two important pathways: endocytosis and exocytosis pathways and their linked to cell signalling, immunity and host-pathogen interactions.

Endocytosis is an essential process used to internalize a wide range of molecules, and involved in many processes such as cell migration, cell division, immunity and pathogen invasion.

- Nathalie identified a family of receptors of cytokines like interleukin-2 receptor (IL-2R) as the first physiological examples of cargos taken up by clathrin independent endocytosis (CIE) (17). They are attractive cargos since they transduce the signal essential for the immune response (18).

- Nathalie’s team further characterized this mechanism at the molecular level and identified a common core complex composed of dynamin, cortactin, Arp2/3 complex, NWASP and F-actin involved in both clathrin-dependent and -independent endocytosis. In addition, specific actors of CIE were uncovered like Rac1, PAKs, the WAVE complex, phosphatidylinositol 3-kinase (PI3K), Vav2 and endophilin (2, 4, 5, 8, 9, 12, 15, 16).

- These results pointed out a key role of PI3K in CIE of cytokine receptors showing for the first time a link between IL-2 signalling and endocytosis (9).

- Moreover, Nathalie’s team reveals a unique mechanism of endocytic pit initiation that does not depend on the deformation of flat membrane, but rather on that of membrane protrusions, due to the specific interaction of the receptor with the WAVE complex (2).

- Finally, this molecular characterization of CIE enabled the recent identification of a growing list of cargoes using this pathway such as α2a- and β1-adrenergic, dopaminergic D3 and D4 receptors, muscarinic acetylcholine receptor 4, EGFR, HGFR, VEGFR, PDGFR, NGFR and IGF1R. Therefore, the mechanism of cytokine receptors endocytosis is a general pathway (4).

In eukaryotic cells, exocytosis is the opposite mechanism than endocytosis. Exocytosis is governed by at least two main compartments, the Golgi complex and recycling endosomes. The distribution of cholesterol in a gradient is essential for exocytosis however the reason why remains obscure. Shigella is an enteroinvasive bacterium that induces bacillary dysentery by invading epithelial cells, causing the destruction of the colon thanks to a T3SS injecting many virulent factors.

- Nathalie’s team has shown that the pathogen Shigella recruits cholesterol at its site of entry in the plasma membrane and once inside, the bacterium impairs the normal distributionof this lipid (11).

- Moreover, Shigella disorganized the Golgi complex and the recycling endosomes, leading to the inhibition of secretion and recycling in infected cells and in animals. Altogether, this effect of Shigella on the cholesterol distribution, on the secretory and recycling networks might have severe consequences on the epithelium barrier function and on the innate immune response (11).

- Recently, Nathalie starts to work on "gut-on-a-chip" technology to investigate host-pathogen interactions.

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