Mapping of the interaction domains of the Crimean-Congo hemorrhagic fever virus nucleocapsid protein.
Crimean-Congo hemorrhagic fever virus (CCHFV) is a member of the genus Nairovirus of the family Bunyaviridae, that can cause severe haemorrhagic fever in humans, with mortality rates above 30  %. CCHFV is the most widespread of the tick-borne human viruses and it is endemic in areas of central Asia, the Middle East, Africa and southern Europe. Its viral genome consists of three negative-sense RNA segments. The large segment (L) encodes a viral RNA-dependent RNA polymerase (L protein), the small segment (S) encodes the nucleocapsid protein (N protein) and the medium segment (M) encodes the envelope proteins. The N protein of bunyaviruses binds genomic RNA, forming the viral ribonucleoprotein (RNP) complex. The L protein interacts with these RNP structures, allowing the initiation of viral replication. The N protein also interacts with actin, although the regions and specific residues involved in these interactions have not yet been described. Here, by means of immunoprecipitation and immunofluorescence assays, we identified the regions within the CCHFV N protein implicated in homo-oligomerization and actin binding. We describe the interaction of the N protein with the CCHFV L protein, and identify the N- and C-terminal regions within the L protein that might be necessary for the formation of these N-L protein complexes. These results may guide the development of potent inhibitors of these complexes that could potentially block CCHFV replication.
Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.|Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Jesica M Levingston
JML
Macleod
Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.|Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Hannah
H
Marmor
Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, NY, USA.|Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.|Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Adolfo
A
García-Sastre
Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, USA.|Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Natalia
N
Frias-Staheli
D015967:Gene Expression Regulation, Viral / Q000502:physiology; D016679:Genome, Viral; D057809:HEK293 Cells; D003225:Hemorrhagic Fever Virus, Crimean-Congo / Q000235:genetics*; D006801:Humans; D019590:Nucleocapsid Proteins / Q000235:genetics*; D011485:Protein Binding; D011487:Protein Conformation; D014779:Virus Replication
D019590:Nucleocapsid Proteins