Systems Biology Reveals NS4B-Cyclophilin A Interaction: A New Target to Inhibit YFV Replication.
Yellow fever virus (YFV) replication is highly dependent on host cell factors. YFV NS4B is reported to be involved in viral replication and immune evasion. Here interactions between NS4B and human proteins were determined using a GST pull-down assay and analyzed using 1-DE and LC-MS/MS. We present a total of 207 proteins confirmed using Scaffold 3 Software. Cyclophilin A (CypA), a protein that has been shown to be necessary for the positive regulation of flavivirus replication, was identified as a possible NS4B partner. 59 proteins were found to be significantly increased when compared with a negative control, and CypA exhibited the greatest difference, with a 22-fold change. Fisher's exact test was significant for 58 proteins, and the p value of CypA was the most significant (0.000000019). The Ingenuity Systems software identified 16 pathways, and this analysis indicated sirolimus, an mTOR pathway inhibitor, as a potential inhibitor of CypA. Immunofluorescence and viral plaque assays showed a significant reduction in YFV replication using sirolimus and cyclosporine A (CsA) as inhibitors. Furthermore, YFV replication was strongly inhibited in cells treated with both inhibitors using reporter BHK-21-rep-YFV17D-LucNeoIres cells. Taken together, these data suggest that CypA-NS4B interaction regulates YFV replication. Finally, we present the first evidence that YFV inhibition may depend on NS4B-CypA interaction.
Laboratório de Virologia, Faculdade de Medicina de José do Rio Preto , São José do Rio Preto, São Paulo 15090-000, Brazil.
Alessandra
A
Vidotto
Laboratório de Virologia, Faculdade de Medicina de José do Rio Preto , São José do Rio Preto, São Paulo 15090-000, Brazil.
Ana T S
AT
Morais
Laboratório de Virologia, Faculdade de Medicina de José do Rio Preto , São José do Rio Preto, São Paulo 15090-000, Brazil.
Milene R
MR
Ribeiro
Laboratório de Virologia, Faculdade de Medicina de José do Rio Preto , São José do Rio Preto, São Paulo 15090-000, Brazil.
Carolina C
CC
Pacca
Laboratório de Virologia, Faculdade de Medicina de José do Rio Preto , São José do Rio Preto, São Paulo 15090-000, Brazil.
Ana C B
AC
Terzian
Departamento de Virologia, Centro de Pesquisa Aggeu Magalhães , Fundação Oswaldo Cruz (FIOCRUZ) - Recife, Pernambuco 50740-465, Brazil.
Laura H V G
LH
Gil
Laboratório de Genômica Estrutural, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro - UFRJ , Rio de Janeiro RJ 21941-902, Brazil.
Ronaldo
R
Mohana-Borges
Department of Immunology & Microbial Science, The Scripps Research Institute - La Jolla , San Diego, California 92037, United States.
Philippe
P
Gallay
Laboratório de Virologia, Faculdade de Medicina de José do Rio Preto , São José do Rio Preto, São Paulo 15090-000, Brazil.
Mauricio L
0000-0003-1102-2419
ML
Nogueira
D021984:Cyclophilin A / Q000235:genetics / Q000378:metabolism*; D054884:Host-Pathogen Interactions / Q000187:drug effects; D006801:Humans; D011506:Proteins / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects; D020123:Sirolimus / Q000008:administration & dosage; D049490:Systems Biology; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D017361:Viral Nonstructural Proteins / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics*; D015005:Yellow fever virus / Q000235:genetics* / Q000472:pathogenicity
C080106:NS4A protein, flavivirus; D011506:Proteins; D017361:Viral Nonstructural Proteins; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D021984:Cyclophilin A; D020123:Sirolimus
NS4B; cyclophilin A; cyclosporine A; protein interactions; proteomics; sirolimus; systems biology; yellow fever virus