Henipaviruses are the newest genus of paramyxoviruses, and recent studies have shown that bats, on a global scale, are host to major genera of mammalian paramyxoviruses3. Paramyxoviruses are membrane enveloped, singled-stranded, negative-sense RNA viruses that include pathogens of global biomedical and agricultural import.

Although henipavirus spillover events are currently limited to Southeast Asia and Australia, a recent global survey of almost 5,000 bat specimens revealed at least 23 distinct viral clades within the henipavirus genus, with the known Nipah and Hendra viruses representing only two of those clades3. The phylogenetic diversity of henipaviruses is thus vastly greater than what was thought previously. Henipaviruses also exhibit a much broader species tropism than other paramyxoviruses, largely due to the highly conserved protein receptors that these viruses use. In the various taxonomic schemes proposed for the transitional dynamics of zoonotic pathogens4,5 all these features justifiably place henipavirus at or close to the penultimate stage for sustained transmission in human outbreaks.

Our laboratory6, and others7, independently discovered EphrinB2 as the major receptor for Nipah and Hendra virus entry. We also identified EphrinB3 as an alternative receptor for henipavirus entry8. EphrinB2 and B3 belong to a class of highly conserved receptor tyrosine kinases (RTKs) that are involved in germ layer differentiation, tissue boundary formation, and other critical developmental processes such as angiogenesis, neurogenesis and axonal guidance9.

Paramyxovirus entry and fusion is a molecular choreography requiring cognate interactions between protein partners, the fusion (F) and attachment (HN/H/G) envelope glycoproteins, in which motion and form must both, be carefully regulated. Please see “Modes of paramyxovirus fusion: a Henipavirus perspective” for a recent review on paramyxovirus fusion from this lab10.

Our research on henipavirus entry have focused on:

  • Elucidating the molecular determinants of attachment protein (G)-receptor specificity2,6,8,11-15
  • Identifying the parameters that modulate fusion (F) protein activity16-18
  • Dissecting the receptor binding triggered membrane fusion cascade18-22
  • Developing quantitative and immunological tools to shed light on the kinetics and stoichiometry of the fusion process14,18,20,23
  • Characterizing the prevalence of cross-reactive neutralizing antibodies from wildlife (mostly bats) and humans at high-risk for exposure in various locales (West Africa, South America) that have now reported the presence of henipavirus-related viruses24

We have developed a highly efficient reverse genetics rescue systems for single-stranded negative-sense RNA viruses (Mononegavirales) that are more robust (easy to optimize) and do not required the use of vaccinia-driven T7 polymerase (a standard method in the field). For full protocol, see Beaty, S.M. et al., mSphere, 2017.


  1. Rissanen, I. et al. Idiosyncratic Mojiang virus attachment glycoprotein directs a host-cell entry pathway distinct from genetically related henipaviruses. Nature Communications 8, 16060, doi: 10.1038/ncomms16060 (2017).
  2. Lee, B. et al. Molecular recognition of human ephrinB2 cell surface receptor by an emergent African henipavirus. Proc Natl Acad Sci U S A 112, E2156-E2165, doi: 10.1073/pnas.1501690112 (2015).
  3. Drexler, J. F. et al. Bats host major mammalian paramyxoviruses. Nature communications 3, 796, doi:10.1038/ncomms1796 (2012).
  4. Wolfe, N. D., Dunavan, C. P. & Diamond, J. Origins of major human infectious diseases. Nature 447, 279-283, doi:10.1038/nature05775 (2007).
  5. Lloyd-Smith, J. O. et al. Epidemic dynamics at the human-animal interface. Science 326, 1362-1367, doi:10.1126/science.1177345 (2009).
  6. Negrete, O. A. et al. EphrinB2 is the entry receptor for Nipah virus, an emergent deadly paramyxovirus. Nature 436, 401-405, doi:10.1038/nature03838 (2005).
  7. Bonaparte, M. I. et al. Ephrin-B2 ligand is a functional receptor for Hendra virus and Nipah virus. Proc Natl Acad Sci U S A 102, 10652-10657, doi:10.1073/pnas.0504887102 (2005).
  8. Negrete, O. A. et al. Two key residues in ephrinB3 are critical for its use as an alternative receptor for Nipah virus. PLoS Pathog 2, e7, doi:10.1371/journal.ppat.0020007 (2006).
  9. Pasquale, E. B. Eph-ephrin bidirectional signaling in physiology and disease. Cell 133, 38-52, doi:10.1016/j.cell.2008.03.011 (2008).
  10. Lee, B. & Ataman, Z. A. Modes of paramyxovirus fusion: a Henipavirus perspective. Trends Microbiol, doi:S0966-842X(11)00053-9 [pii] 10.1016/j.tim.2011.03.005 (2011).
  11. Lee, B. Envelope-receptor interactions in Nipah virus pathobiology. Ann N Y Acad Sci 1102, 51-65, doi:1102/1/51 [pii] 10.1196/annals.1408.004 (2007).
  12. Negrete, O. A., Chu, D., Aguilar, H. C. & Lee, B. Single amino acid changes in the Nipah and Hendra virus attachment glycoproteins distinguish ephrinB2 from ephrinB3 usage. J Virol 81, 10804-10814, doi:JVI.00999-07 [pii] 10.1128/JVI.00999-07 (2007).
  13. Lee, B., Ataman, Z. A. & Jin, L. Evil versus ‘eph-ective’ use of ephrin-B2. Nat Struct Mol Biol 15, 540-542, doi:nsmb0608-540 [pii] 10.1038/nsmb0608-540 (2008).
  14. Wolf, M. C. et al. A catalytically and genetically optimized beta-lactamase-matrix based assay for sensitive, specific, and higher throughput analysis of native henipavirus entry characteristics. Virol J 6, 119, doi:10.1186/1743-422x-6-119 (2009).
  15. Pernet, O., Wang, Y. E. & Lee, B. Henipavirus Receptor Usage and Tropism. Current topics in microbiology and immunology, doi:10.1007/82_2012_222 (2012).
  16. Aguilar, H. C. et al. N-glycans on Nipah virus fusion protein protect against neutralization but reduce membrane fusion and viral entry. J Virol 80, 4878-4889, doi:10.1128/jvi.80.10.4878-4889.2006 (2006).
  17. Aguilar, H. C. et al. Polybasic KKR motif in the cytoplasmic tail of Nipah virus fusion protein modulates membrane fusion by inside-out signaling. J Virol 81, 4520-4532, doi:10.1128/jvi.02205-06 (2007).
  18. Aguilar, H. C., Aspericueta, V., Robinson, L. R., Aanensen, K. E. & Lee, B. A quantitative and kinetic fusion protein-triggering assay can discern distinct steps in the nipah virus membrane fusion cascade. J Virol 84, 8033-8041, doi:10.1128/jvi.00469-10 (2010).
  19. Aguilar, H. C. et al. A novel receptor-induced activation site in the Nipah virus attachment glycoprotein (G) involved in triggering the fusion glycoprotein (F). J Biol Chem 284, 1628-1635, doi:M807469200 [pii] 10.1074/jbc.M807469200 (2009).
  20. Garner, O. B. et al. Endothelial galectin-1 binds to specific glycans on nipah virus fusion protein and inhibits maturation, mobility, and function to block syncytia formation. PLoS Pathog 6, e1000993, doi:10.1371/journal.ppat.1000993 (2010).
  21. Wolf, M. C. et al. A broad-spectrum antiviral targeting entry of enveloped viruses. Proc Natl Acad Sci U S A 107, 3157-3162, doi:10.1073/pnas.0909587107 (2010).
  22. Mirza, A. M. et al. Triggering of the newcastle disease virus fusion protein by a chimeric attachment protein that binds to Nipah virus receptors. J Biol Chem 286, 17851-17860, doi:M111.233965 [pii] 10.1074/jbc.M111.233965 (2011).
  23. Tamin, A. et al. Development of a neutralization assay for Nipah virus using pseudotype particles. J Virol Methods 160, 1-6, doi:S0166-0934(09)00103-7 [pii] 10.1016/j.jviromet.2009.02.025 (2009).
  24. Pernet, O. et al. Evidence for henipavirus spillover into human populations in Africa. Nature communications 5, 5342, doi: 10.1038/ncomms6342 (2014).