Structure and function of the Toscana virus cap-snatching endonuclease.
Biocatalysis
Catalytic Domain
Cations, Divalent
/ pharmacology
Conserved Sequence
DNA Replication
/ drug effects
Endonucleases
/ chemistry
Enzyme Inhibitors
/ pharmacology
Enzyme Stability
/ drug effects
Models, Molecular
Mutant Proteins
/ metabolism
Protein Domains
RNA Caps
/ metabolism
Sandfly fever Naples virus
/ enzymology
Static Electricity
Sulfates
/ metabolism
Transcription, Genetic
/ drug effects
Viral Proteins
/ chemistry
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
18 11 2019
18 11 2019
Historique:
accepted:
01
10
2019
revised:
14
09
2019
received:
11
03
2019
pubmed:
5
10
2019
medline:
13
5
2020
entrez:
5
10
2019
Statut:
ppublish
Résumé
Toscana virus (TOSV) is an arthropod-borne human pathogen responsible for seasonal outbreaks of fever and meningoencephalitis in the Mediterranean basin. TOSV is a segmented negative-strand RNA virus (sNSV) that belongs to the genus phlebovirus (family Phenuiviridae, order Bunyavirales), encompassing other important human pathogens such as Rift Valley fever virus (RVFV). Here, we carried out a structural and functional characterization of the TOSV cap-snatching endonuclease, an N terminal domain of the viral polymerase (L protein) that provides capped 3'OH primers for transcription. We report TOSV endonuclease crystal structures in the apo form, in complex with a di-ketoacid inhibitor (DPBA) and in an intermediate state of inhibitor release, showing details on substrate binding and active site dynamics. The structure reveals substantial folding rearrangements absent in previously reported cap-snatching endonucleases. These include the relocation of the N terminus and the appearance of new structural motifs important for transcription and replication. The enzyme shows high activity rates comparable to other His+ cap-snatching endonucleases. Moreover, the activity is dependent on conserved residues involved in metal ion and substrate binding. Altogether, these results bring new light on the structure and function of cap-snatching endonucleases and pave the way for the development of specific and broad-spectrum antivirals.
Identifiants
pubmed: 31584100
pii: 5580917
doi: 10.1093/nar/gkz838
pmc: PMC6847833
doi:
Substances chimiques
Cations, Divalent
0
Enzyme Inhibitors
0
Mutant Proteins
0
RNA Caps
0
Sulfates
0
Viral Proteins
0
Endonucleases
EC 3.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
10914-10930Informations de copyright
© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.
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