Intracellular transport of herpes simplex virus type 1 (HSV-1) capsids

The long term objectives of the laboratory are to characterize host-pathogen interactions. Our work focuses on the molecular characterization of the intracellular pathway taken by the HSV-1 capsids by defining the molecules involved in the process. Newly assembled viral capsids are assembled in the nucleus, but these viral particles are too big to reach the cytoplasm through the nuclear pores. The capsids consequently uses a unique mean to traverse the two nuclear membranes by budding through the inner nuclear membrane and then rapidly fusing with the outer nuclear membrane to release unenveloped capsids in the cytoplasm (HSV-1 nuclear exit model). To study this process, our lab has developed a world’s unique in vitro assay that reconstitutes nuclear egress in the test tube. That assay has enables us to clarify how the virus escapes the nucleus and resulted in the identification, by mass spectrometry, of novel components of the nuclear capsids. In addition, we are studying the targeting of viral glycoproteins at the nuclear membranes and probing their role there. Among them, we showed that the viral glycoprotein M is rapidly and specifically targeted to the nucleus well before the virus leaves that compartment. However, its implication in viral egress is not clear. We do know however that it binds to many other proteins whose functions we are examining.

Another critical step in the egress of the newly produced viral particles is their acquisition of a final and mature envelope. Work by our laboratory and many others suggests the implication of the TGN, albeit the mechanism of this process remains largely mysterious for the moment. Finally, the virus must then escape the TGN to reach the cell surface. Our lab has demonstrated that the virus reintegrates the classical biosynthetic pathway and we identified one important regulator of that route, namely the host protein kinase D.

The virus thus follows a highly complex pathway to release its newly formed viral particles. Unfortunately, little is known of the molecular details of this pathway as well as the contribution of the cell to this process. To better understand this, we put together a proteomics program to analyze the compositions of viral intermediates and reported 49 host proteins on mature virions, many of which proved to regulate the viral life cycle as revealed in a siRNA screen. We are currently characterizing the role of these proteins. In parallel, we are following the maturation of the virus by flow virometry, a highly innovative approach we developed and allows us to monitor and even purify individual viral particles.

Available lab positions 

Master, doctorate or postdoc:

Bilali, N.¹, Khadivjam, B.¹, Bonneil, E., Thibault, P. and Lippé R. (2020). Proteomics of Herpes simplex virus type 1 nuclear capsids. J Virol (in press)              NB:  The first two authors equally contributed

Boruchowicz, H, Hawkins, J, Cruz-Palomar, K and Lippé, R. (2020). The XPO6 exporting mediates HSV-1 gM nuclear release late in infection. J Virol Online ahead of print

Lippé, R. (2020). Intracellular transport of Alphaherpesvirinae. Virologie 24:210-30

Khadivjam, B., El Bilili, N. and Lippé, R. (2020). Analysis and sorting of individual viral particles by flow virometry. Methods Mol Biol. 2060: 289-303

Roussel É and Lippé R. (2018). Cellular Protein Kinase D Modulators Play a Role during Multiple Steps of Herpes Simplex Virus Type 1 Egress. J Virol. 92: e01486-18

Lippé, R. (2018). Flow Virometry: a Powerful Tool To Functionally Characterize Viruses. J Virol 92: e01765-17

El Kasmi I, Khadivjam B, Lackman M, Duron J, Bonneil E, Thibault  and Lippé R. (2017). Extended synaptotagmin 1 interacts with the Herpes simplex virus type 1 glycoprotein M and negatively modulates virus-induced membrane fusion. J Virol 92: e01281-17

Rodriguez, L., Nguyen-Vi, M., Desjardins, A., Lippé, R., Fon, E. and Leclerc N. (2017). Rab7a regulates Tau secretion. J Neurochemistry 141: 592-605

Striebinger H, Zhang J, Ott M, Funk C, Radtke K, Duron J, Ruzsics Z, Haas J, Lippé R, Bailer SM. (2015). Subcellular trafficking and functional importance of Herpes simplex virus type 1 Glycoprotein M domains. J Gen Virol. 96(11):3313-25.

El Kasmi I, Lippé R. (2015). Herpes simplex virus 1 gN partners with gM to modulate the viral fusion machinery. J Virol. 89(4):2313-23.

Lippé, R (2014). Characterization of extracellular HSV-1 virions by proteomics. Methods Mol Biol. 1144:181-90.

Henaff, D., Rémillard-Labrosse, G., Loret, S. and Lippé, R. (2013). Analysis of the early steps of herpes simplex virus type 1 capsid tegumentation. J Virol 87:4895-4906.

Stegen, C., Yakova, Y., Henaff, D., Nadjar, J., Duron, J. and Lippé,  R. (2013). Analysis of Virion-Incorporated Host Proteins Required for Herpes Simplex Virus Type 1 Infection through a RNA Interference Screen. PLOS ONE 8:e53276.

Radtke, K., English, L., Rondeau, C., Leib, D., Lippé, R. and Desjardins, M. (2013). Inhibition of the Host Translation Shut-off Response by Herpes Simplex Virus 1 triggers Nuclear Envelope-derived Autophagy. J Virol 87:3990-7. (JVI Spotlight*⁵)

Henaff, D., Radtke, K. and Lippé, R. (2012). Herpesviruses exploit several host compartments for envelopment. Traffic 13(11):1443-9.

Loret, S. El Bilali, N. and Lippé, R. (2012). Analysis of herpes simplex virus type I nuclear particles by flow cytometry. Cytometry Part A 81A: 950–959. (see Commentary by Dorothy E Lewis as well as In This Issue Feature*)

Lippé, R. (2012). Deciphering novel host-herpesvirus interactions by proteomics. Frontiers Microbiol 181:1-14.

Loret S. and Lippé, R. (2012). Biochemical Analysis of ICP0, ICP4, UL7 and UL23 Incorporated Into Extracellular Herpes Simplex Type 1 Virions. J Gen Virol 93:624-34.

Rémillard-Labrosse, G. and Lippé, R. (2011). In vitro nuclear egress of herpes simplex virus type 1 capsids. Methods 55: 153-159.

Zhang J, Nagel CH, Sodeik B and Lippé R. (2009). Early, active and specific localization of HSV-1 gM to nuclear membranes. J Virol 83:12984-12997.

Rémillard-Labrosse, G., Mihai, C., Duron, J., Guay, G. and Lippé, R. (2009). Protein kinase D dependent trafficking of the large Herpes simplex virus type 1 capsids from the TGN to plasma membrane. Traffic 10:1074-1083.

Rémillard-Labrosse, G. and Lippé, R. (2009). Meeting of conventional and unconventional pathways at the TGN. Commun Integr Biol 2: 434-436.

English, L., Chemali, M., Duron, J., Rondeau, C., Laplante, A., Gingras, D., Alexander, D., Leib, D., Norbury, C., Lippé, R. and Desjardins, M. (2009). Autophagy enhances the presentation of endogenous viral antigens on MHC class I molecules during HSV-1 infection. Nature Immunol. 5:480-7.

Loret, S., Guay, G. and Lippé, R. (2008). Comprehensive characterization of extracellular HSV-1 virions. J Virol. 82:8605-18. (Cover page**)

Rémillard-Labrosse, G. Guay, G. and Lippé, R. (2006). Reconstitution of HSV-1 nuclear capsid egress in vitro. J Virol. 80:9741-53. (JVI Spotlight***)

Turcotte, S. Letellier, J. and Lippé, R. (2005). HSV-1 capsids transit by the TGN, where viral glycoproteins accumulate independently of capsid egress. J Virol. 79: 8847-60. (JVI Spotlight*⁴)

Moise, A.R., Grant J.R., Lippé R., Gabathuler, R. and Jefferies, W.A. (2004). The adenovirus E3-6.7K protein adopts diverse membrane topologies following posttranslational translocation. J Virol. 78: 454-63.

Lippé, R., Miaczynska, M, Rybin, V., Runge, A. and Zerial, M. (2001). Functional synergy between the Rab5 effector Rabaptin-5 and the exchange factor Rabex-5 when physically associated in a complex. Mol. Biol. Cell, 12:2219-2228 .

Lippé, R., Horiuchi, H. and Zerial, M. (2001). Expression, purification and characterization of the Rab5 effector complex, Rabaptin-5/Rabex-5 in Regulators and effectors of small GTPases.  Abelson, J.N. and Simon, M.I. eds.  Methods Enzymol., 329:132-145 .

Rubino, M., Miaczynska M., Lippé, R. and Zerial, M. (2000).  Selective membrane recruitment of EEA1 suggests a role in directional transport of clathrin-coated vesicles to early endosomes.  J.Biol.Chem. 275: 3745-3748.

Simonsen, A.¹ Lippé, R.¹, Christoforidis, S., Gaullier, J.M., Brech, A., Callaghan, J., Toh, B.H., Murphy, C., Zerial, M. and Stenmark, H. (1998). EEA1 links PI(3)K function to Rab5 regulation of endosome fusion.  Nature, 394:494-498.                   NB:  The first two authors equally contributed

Gournier, H., Stenmark, H., Rybin, V., Lippé, R. and Zerial, M. (1998).  Two distinct effectors of the small GTPase Rab5 cooperate in endocytic membrane fusion.  EMBO J. 17:  1930-1940 .

Horiuchi, H., Lippé, R., McBride, H.M., Rubino, M., Woodman, P., Stenmark, H., Rybin, V., Wilm, M., Ashman, K., Mann, M., and Zerial, M.  (1997).  A novel Rab5 GDP/GTP exchange factor complexed to Rabaptin-5 links nucleotide exchange to effector recruitment and function.  Cell 90: 1149-1159.

Lippé, R., Kolaitis, G., Michaelis, C., Tufaro, F., and Jefferies, W.A. (1993).  Differential recruitment of viral and allo-epitopes into the MHC class I antigen processing pathway of novel mutant Ltk- cells.  J. Immunol. 150: 3170-3179.

Michaelis, C., Banfield, B.W., Gruenheid, S., Tsang, Y., Lippé, R., Jefferies, W.A., Wattenberg, B.W., and Tufaro, F. (1992).  Toxin resistance and reduced secretion in a mouse L-cell mutant defective in herpes virus propagation.  Biochem. Cell Biol. 70:1209-1217 .

Lippé, R., Luke,E., Kuah, Y.T., Lomas, C., and Jefferies, W.A. (1991).  Adenovirus infection inhibits the phosphorylation of major histocompatibility complex class I proteins.  J. Exp. Med. 174:1159-1166 .

Lippé, R. and Graham, F.L. (1989).  Adenoviruses with nonidentical terminal sequences are viable.  J. Virol. 63:5133-5141 .

Special mentions

*        Comment by D Lewis (2012). Sweat the small stuff.

          Also see: "In This Issue Feature"  Small is beautiful

**       Made the cover page of J Virol (2008) volume 82 number 20 (mi-octobre)

***     JVI Spotlight: Test Tube Herpesvirus Leaving Its Nuclear Birthplace, J Virol. (2006) 80: 9347–8

*⁴     JVI Spotlight: Close Encounter: HSV-1 Capsids and Glycoproteins Travel Independently and Meet at the Site of  Re-Envelopment, J. Virol. (2005) 79:8675–6

*⁵      JVI Spotlight: Recoiling from the Nucleus: an Alternative Autophagic Pathway for Antiviral Defense, J. Virol (2013) 87:3916

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Lab members

  Actual team                               

    Past "graduates"                                                                                                       

                                                    

                 

                                     

         

                

Past undergraduate trainees

                                                     
         
as well as:
- Mohamed Madi (2009)
- Thimothée Govare (2010)
- Colin Deschanvres (2010)
- Laure Cockenpot (2011)
- Nicolas Bensid (2011)
- Dave Bauer (2013)
- Pierrette Kwemo (2015)

             
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Contact us

                    Office: 6.17.006 (Centre de recherche du CHU Sainte-Justine)

                    Tel: (514) 345-4931 ext 5201

                    Email: roger.lippe@umontreal.ca

          LinkedIn: here

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Lab funding

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Last update: September 9th, 2020