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Hantavirus RdRp Requires a Host Cell Factor for Cap Snatching.

Subbiah Jeeva1, Sheema Mir2, Adrain Velasquez3

  • 1Western University of Health Sciences, Pomona, California, USA.

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|December 14, 2018
PubMed
Summary

Hantavirus uses a cap-snatching mechanism to replicate, hijacking host mRNA. A host cell factor is crucial for this process, offering a potential therapeutic target against hantavirus infections.

Keywords:
cap snatchinghantavirusnegative-strand RNA virusnucleocapsid

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Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Hantavirus replication relies on cap-snatching, where viral RNA-dependent RNA polymerase (RdRp) uses host mRNA fragments as primers.
  • The Hantavirus nucleocapsid (N) protein binds 5' mRNA caps, protecting them and facilitating primer generation.
  • The RdRp's N-terminal endonuclease domain exhibits nonspecific RNA degradation, posing a challenge for understanding specific primer generation.

Purpose of the Study:

  • To elucidate the mechanism of hantavirus cap-snatching and primer generation.
  • To investigate the role of the RdRp's endonuclease domain and N protein interaction.
  • To identify potential host factors involved in hantavirus replication.

Main Methods:

  • Construction and testing of a fused RdRp mutant (NC mutant) combining endonuclease and N protein-binding domains.
  • In vitro assays using purified NC mutant, N protein, and human umbilical vein endothelial cell (HUVEC) lysates.
  • Analysis of specific endonucleolytic cleavage of mRNA to generate capped primers.

Main Results:

  • The NC mutant, with N protein, successfully generated specific capped primers in cells.
  • Purified NC mutant and N protein required HUVEC lysates for specific mRNA cleavage in vitro, producing primers of defined length and 3' terminus.
  • Evidence suggests an unknown host cell factor mediates the interaction between N protein and the NC mutant for precise cleavage.

Conclusions:

  • A host cell factor is essential for the specific interaction between Hantavirus N protein and RdRp's endonuclease domain.
  • This host factor facilitates the precise cleavage of capped mRNA fragments, generating functional primers for viral replication.
  • Understanding this mechanism reveals a potential therapeutic target for hantavirus infections, for which no treatment currently exists.