Structural insights into the multifunctionality of rabies virus P3 protein

Proc Natl Acad Sci U S A. 2023 Apr 4;120(14):e2217066120. doi: 10.1073/pnas.2217066120. Epub 2023 Mar 29.


Viruses form extensive interfaces with host proteins to modulate the biology of the infected cell, frequently via multifunctional viral proteins. These proteins are conventionally considered as assemblies of independent functional modules, where the presence or absence of modules determines the overall composite phenotype. However, this model cannot account for functions observed in specific viral proteins. For example, rabies virus (RABV) P3 protein is a truncated form of the pathogenicity factor P protein, but displays a unique phenotype with functions not seen in longer isoforms, indicating that changes beyond the simple complement of functional modules define the functions of P3. Here, we report structural and cellular analyses of P3 derived from the pathogenic RABV strain Nishigahara (Nish) and an attenuated derivative strain (Ni-CE). We identify a network of intraprotomer interactions involving the globular C-terminal domain and intrinsically disordered regions (IDRs) of the N-terminal region that characterize the fully functional Nish P3 to fluctuate between open and closed states, whereas the defective Ni-CE P3 is predominantly open. This conformational difference appears to be due to the single mutation N226H in Ni-CE P3. We find that Nish P3, but not Ni-CE or N226H P3, undergoes liquid-liquid phase separation and this property correlates with the capacity of P3 to interact with different cellular membrane-less organelles, including those associated with immune evasion and pathogenesis. Our analyses propose that discrete functions of a critical multifunctional viral protein depend on the conformational arrangements of distant individual domains and IDRs, in addition to their independent functions.

Keywords: NNS RNA virus; liquid-liquid phase separation; membrane-less organelles; protein multifunctionality; rabies lyssavirus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Humans
  • Protein Isoforms / metabolism
  • Rabies virus* / genetics
  • Rabies*
  • Viral Proteins / genetics
  • Viral Proteins / metabolism
  • Virulence Factors / metabolism


  • Viral Proteins
  • Virulence Factors
  • Protein Isoforms