domingo, 5 de abril de 2015

Virus-like particle system identifies the endonuclease domain of Crimean-Congo hemorrhagic fever virus. - PubMed - NCBI

Virus-like particle system identifies the endonuclease domain of Crimean-Congo hemorrhagic fever virus. - PubMed - NCBI



 2015 Mar 25. pii: JVI.03691-14. [Epub ahead of print]

Virus-like particle system identifies the endonuclease domain of Crimean-Congo hemorrhagic fever virus.

Abstract

Crimean-Congo Hemorrhagic Fever virus (CCHFV; genus Nairovirus) is an extremely pathogenic member of the Bunyaviridae family. Since handling of the virus requires a BSL-4 facility, little is known about pathomechanisms and host interactions. Here, we describe the establishment of a transcriptionally competent (tc)-VLP system for CCHFV. Recombinant polymerase (L), nucleocapsid protein (N) and a reporter minigenome expressed in human HuH-7 cells resulted in formation of transcriptionally active nucleocapsids that could be packaged by co-expressed CCHFV glycoproteins into tc-VLPs. The tc-VLPs resembled authentic virus particles in their protein composition and neutralization sensitivity to anti-CCHFV antibodies, and could recapitulate all steps of the viral replication cycle. Particle attachment, entry and primary transcription were modelled by infection of naive cells. The subsequent steps of genome replication, secondary transcription, and particle assembly and release can be obtained upon passaging the tc-VLPs on cells expressing CCHFV structural proteins. The utility of the VLP system was demonstrated by showing that the endonuclease domain of L is located around amino acid D693, as it was predicted in silico by Morin et al. (PLoS Pathogens 6: e1001038). The tc-VLP system will greatly facilitate studies and diagnostics of CCHFV under non-BSL-4 conditions.

IMPORTANCE:

Crimean-Congo Hemorrhagic Fever virus (CCHFV) is an extremely virulent pathogen of humans. Since the virus can only be handled at the highest biosafety level, research is restricted to a few specialized laboratories. We developed a plasmid-based system to produce virus-like particles with the ability to infect cells and transcribe a reporter genome. Due to the absence of viral genes, the virus-like particles are unable to spread or cause disease, thus allowing to study aspects of CCHFV biology under relaxed biosafety conditions.
Copyright © 2015, American Society for Microbiology. All Rights Reserved.

PMID:
 
25810550
 
[PubMed - as supplied by publisher]

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