Flavivirus NS3 and NS5 proteins interaction network: a high-throughput yeast two-hybrid screen.
BACKGROUND
The genus Flavivirus encompasses more than 50 distinct species of arthropod-borne viruses, including several major human pathogens, such as West Nile virus, yellow fever virus, Japanese encephalitis virus and the four serotypes of dengue viruses (DENV type 1-4). Each year, flaviviruses cause more than 100 million infections worldwide, some of which lead to life-threatening conditions such as encephalitis or haemorrhagic fever. Among the viral proteins, NS3 and NS5 proteins constitute the major enzymatic components of the viral replication complex and are essential to the flavivirus life cycle.


RESULTS
We report here the results of a high-throughput yeast two-hybrid screen to identify the interactions between human host proteins and the flavivirus NS3 and NS5 proteins. Using our screen results and literature curation, we performed a global analysis of the NS3 and NS5 cellular targets based on functional annotation with the Gene Ontology features. We finally created the first flavivirus NS3 and NS5 proteins interaction network and analysed the topological features of this network. Our proteome mapping screen identified 108 human proteins interacting with NS3 or NS5 proteins or both. The global analysis of the cellular targets revealed the enrichment of host proteins involved in RNA binding, transcription regulation, vesicular transport or innate immune response regulation.


CONCLUSIONS
We proposed that the selective disruption of these newly identified host/virus interactions could represent a novel and attractive therapeutic strategy in treating flavivirus infections. Our virus-host interaction map provides a basis to unravel fundamental processes about flavivirus subversion of the host replication machinery and/or immune defence strategy.
Inserm Unit 851, Lyon, France.
Marc
M
Le Breton
Laurène
L
Meyniel-Schicklin
Alexandre
A
Deloire
Bruno
B
Coutard
Bruno
B
Canard
Xavier
X
de Lamballerie
Patrice
P
Andre
Chantal
C
Rabourdin-Combe
Vincent
V
Lotteau
Nathalie
N
Davoust
D005416:Flavivirus / Q000472:pathogenicity; D018177:Flavivirus Infections / Q000821:virology; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D054884:Host-Pathogen Interactions; D006801:Humans; D025941:Protein Interaction Mapping; D020365:RNA Helicases / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D020365:RNA Helicases; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C096041:NS5 protein, flavivirus