In:
The Journal of Immunology, The American Association of Immunologists, Vol. 198, No. 1_Supplement ( 2017-05-01), p. 214.8-214.8
Abstract:
Ebola virus causes severe hemorrhagic fevers with high mortality characterized by unchecked viral growth and inflammation with marked suppression of host innate and adaptive immune responses. Ebola virus infects macrophages and dendritic cells preferentially, repressing the development of MHC and antigen presentation. Patients who succumb to Ebola virus disease show little evidence of a humoral immune response. VP35 inhibits critical immune signaling early in infection. We have carried out global phosphoproteomic analysis of Ebola virions produced in monkey and bat cells using high resolution MS/MS. We detected all viral proteins with high coverage and identified novel phosphorylation sites in all seven proteins of each virion type. We compared phosphorylation patterns and validated the results by testing the role of the phosphorylation sites of interferon-modulatory VP35 using mutagenesis, in vivo phosphorylation and minigenome assay. We carried out biophysical analyses, structural modeling and simulations of wildtype versus phosphorylated proteins. The phosphorylation pattern differs between virions grown in monkey cells and those grown in bat cells. Nearly all phosphorylation sites were found to be solvent accessible with the majority in flexible regions. VP30, VP35 and NP were characterized by inherent disorder. Mutagenesis, phosphorylation analysis and minigenome experiments showed that VP35 Ser-210 undergoes phosphorylation and is important for transcription. Ser-210 lies in a linker region connecting the coiled coil oligomerization domain and the interferon inhibitory domain. Our findings point to an important role of VP35 phosphorylation and a means of developing inhibitors as drugs against Ebola virus.
Type of Medium:
Online Resource
ISSN:
0022-1767
,
1550-6606
DOI:
10.4049/jimmunol.198.Supp.214.8
Language:
English
Publisher:
The American Association of Immunologists
Publication Date:
2017
detail.hit.zdb_id:
1475085-5
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