The role of host cell proteins in the transcription and replication of Ebola virus
Jordana is a BSc graduate in Microbiology and has an MSc in Medical Research. Her interest in molecular biology, infectious diseases and enthusiasm to live in another country made her pack her bags and relocate to Liverpool to join the HPRU EZI and Julian Hiscox’s group in order to start a PhD on ‘The role of host cell proteins in the transcription and replication of Ebola virus’.
She loves both reading a good book with a nice mug of coffee and hanging out with her friends in the vibrant city of Liverpool.
The Ebola virus (EBOV) ribonucleoprotein complex (RNP) proteins are essential for the viral genome transcription and replication. The viral RNA-dependent RNA polymerase (L) plays a major role in these processes and thus represents an attractive therapeutic target. VP35, its co-factor, is also involved in the antagonism of the host interferon (IFN) response.
Label-free proteomics are a useful tool to identify and quantify the host proteins that potentially interact with the viral ones. In this project, both Ebola virus L and VP35 cellular interactomes have been elucidated by LC-MS/MS. Calmodulin (CALM) and the Cytoplasmic Dynein light chain 1 (DYNLL1) were identified as significant VP35 and L interactors, respectively. When knocking them down with the CALM antagonist W-7 and the DYNLL1 antagonist Ciliobrevin D in mammalian cell cultures were a minigenome system is being used as a model for EBOV transcription and replication, the reporter activity of it droped down in a dose-response manner. These findings suggested those two cellular proteins do have a role in the viral processes.
In the second part of the project, three non-synonymous mutations found to have occurred in EBOV VP35 during the 2014-2016 EBOV West Africa outbreak will be studied. 950 VP35 amino acid sequences downloaded from GenBank were compared to an EBOV reference sequence (NCBI AGB56837.1) and T68M, L249F and L330F were the most occurring ones. A minigenome system will assess whether these mutations do have an effect on the viral transcription and replication, and an IFN assay will determine if T68M, L249F and L330F are determinants of the role in IFN antagonism of VP35.
- Julian A. Hiscox - NIHR Health Protection Research Unit in Emerging and Zoonotic Infections, University of Liverpool
- Miles W. Carroll - NIHR Health Protection Research Unit in Emerging and Zoonotic Infections, Public Health England