Biophysical and structural characterization of the recombinant human eIF3L
| Full text | |
| Author(s): |
Vidotto, Alessandra
;
Morais, Ana T. S.
;
Ribeiro, Milene R.
;
Pacca, Carolina C.
;
Terzian, Ana C. B.
;
Gil, Laura H. V. G.
;
Mohana-Borges, Ronaldo
;
Gallay, Philippe
;
Nogueira, Mauricio L.
Total Authors: 9
|
| Document type: | Journal article |
| Source: | JOURNAL OF PROTEOME RESEARCH; v. 16, n. 4, p. 1542-1555, APR 2017. |
| Web of Science Citations: | 6 |
| Abstract | |
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 treated with both inhibitors using reporter BHK-21-rep-YFV17DLucNeolres 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. (AU) | |
| FAPESP's process: | 09/01400-7 - Study of the interaction of NS4B of yellow fever virus and host cellular proteins |
| Grantee: | Maurício Lacerda Nogueira |
| Support type: | Regular Research Grants |
| FAPESP's process: | 10/05043-1 - Characterization of the interaction of the NS5 protein of Yellow Fever with the human cellular protein eIF3L |
| Grantee: | Ana Theresa Silveira de Morais |
| Support type: | Scholarships in Brazil - Doctorate (Direct) |