title: Characterization of the Interaction between hantavirus nucleocapsid protein (N) and ribosomal protein S19 (RPS19).
abstract: Hantaviruses, members of the Bunyaviridae family, are negative-stranded emerging RNA viruses and category A pathogens that cause serious illness when transmitted to humans through aerosolized excreta of infected rodent hosts. Hantaviruses have evolved a novel translation initiation mechanism, operated by nucleocapsid protein (N), which preferentially facilitates the translation of viral mRNAs. N binds to the ribosomal protein S19 (RPS19), a structural component of the 40 S ribosomal subunit. In addition, N also binds to both the viral mRNA 5' cap and a highly conserved triplet repeat sequence of the viral mRNA 5' UTR. The simultaneous binding of N at both the terminal cap and the 5' UTR favors ribosome loading on viral transcripts during translation initiation. We characterized the binding between N and RPS19 and demonstrate the role of the N-RPS19 interaction in N-mediated translation initiation mechanism. We show that N specifically binds to RPS19 with high affinity and a binding stoichiometry of 1:1. The N-RPS19 interaction is an enthalpy-driven process. RPS19 undergoes a conformational change after binding to N. Using T7 RNA polymerase, we synthesized the hantavirus S segment mRNA, which matches the transcript generated by the viral RNA-dependent RNA polymerase in cells. We show that the N-RPS19 interaction plays a critical role in the translation of this mRNA both in cells and rabbit reticulocyte lysates. Our results demonstrate that the N-mediated translation initiation mechanism, which lures the host translation machinery for the preferential translation of viral transcripts, primarily depends on the N-RPS19 interaction. We suggest that the N-RPS19 interaction is a novel target to shut down the N-mediated translation strategy and hence virus replication in cells.
authors.affiliation: Department of Microbiology, Molecular Genetics, and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66103, USA.
authors.forename: Erdong
authors.initials: E
authors.lastname: Cheng
authors.forename: Absarul
authors.initials: A
authors.lastname: Haque
authors.forename: Mary Ashley
authors.initials: MA
authors.lastname: Rimmer
authors.forename: Islam T M
authors.initials: IT
authors.lastname: Hussein
authors.forename: Sheema
authors.initials: S
authors.lastname: Sheema
authors.forename: Alex
authors.initials: A
authors.lastname: Little
authors.forename: Mohammad A
authors.initials: MA
authors.lastname: Mir
mesh_terms: D020121:5' Untranslated Regions / Q000502:physiology; D000818:Animals; D006476:Orthohantavirus / Q000502:physiology*; D018778:Hantavirus Infections / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D006367:HeLa Cells; D006801:Humans; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D010442:Peptide Chain Initiation, Translational / Q000502:physiology*; D011485:Protein Binding; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*; D054682:Ribosome Subunits, Small, Eukaryotic / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000502:physiology*
chemical_list: D020121:5' Untranslated Regions; D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D012269:Ribosomal Proteins; C040343:ribosomal protein S19; D012324:RNA-Dependent RNA Polymerase