VP30 is a phosphoprotein essential for the initiation of Ebola virus transcription. In this work, we have studied the effect
of mutations in VP30 phosphorylation sites on the ebolavirus replication cycle by using a reverse genetics system. We demonstrate
that VP30 is involved in reinitiation of gene transcription and that this activity is affected by mutations at the phosphorylation
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"This mutation separates clade 2 from clade 3, which carries the derived allele and covers 56% of the sequences. VP30 is a virus-specific transcription factor that mediates transcription activation , while VP24 suppresses interferon production  and also inhibits viral transcription and replication . All other mutations occur on peripheral branches of the tree and have derived allele frequencies below 8%. "
[Show abstract][Hide abstract] ABSTRACT: The 2014 epidemic of the Ebola virus is governed by a genetically diverse
viral population. In the early Sierra Leone outbreak, a recent study has
identified new mutations that generate genetically distinct sequence clades.
Here we find evidence that major Sierra Leone clades have systematic
differences in growth rate and reproduction number. If this growth
heterogeneity remains stable, it will generate major shifts in clade
frequencies and influence the overall epidemic dynamics on time scales within
the current outbreak. Our method is based on simple summary statistics of clade
growth, which can be inferred from genealogical trees with an underlying
clade-specific birth-death model of the infection dynamics. This method can be
used to perform realtime tracking of an evolving epidemic and identify emerging
clades of epidemiological or evolutionary significance.
"Minigenome assays revealed that NP, VP35, and L are essential and sufficient to support viral replication. For efficient transcription, the transcription factor VP30 is also required (Enterlein et al., 2006; Groseth et al., 2005; Martinez et al., 2008, 2011; Mühlberger et al., 1999; Watanabe et al., 2004). The polymerase cofactor VP35 is the functional equivalent of the phosphoprotein P of other NNS RNA viruses (Mühlberger et al., 1999). "
[Show abstract][Hide abstract] ABSTRACT: The Ebola virus (EBOV) RNA-dependent RNA polymerase (RdRp) complex consists of the catalytic subunit of the polymerase, L, and its cofactor VP35. Using immunofluorescence analysis and coimmunoprecipitation assays, we mapped the VP35 binding site on L. A core binding domain spanning amino acids 280-370 of L was sufficient to mediate weak interaction with VP35, while the entire N-terminus up to amino acid 380 was required for strong VP35-L binding. Interestingly, the VP35 binding site overlaps with an N-terminal L homo-oligomerization domain in a non-competitive manner. N-terminal L deletion mutants containing the VP35 binding site were able to efficiently block EBOV replication and transcription in a minigenome system suggesting the VP35 binding site on L as a potential target for the development of antivirals.
"While the kinetics of VP30 expression is tightly regulated in wild-type EBOV-infected cells, VP30 is constantly over-expressed in the EbolaΔVP30 system. Because viral replication and transcription are dependent on the expression level of VP3042325263536373839, the kinetics of viral genome transcription, replication, and gene expression may differ between EbolaΔVP30 and wild-type EBOV. Therefore, some of the findings described in this study should be repeated with authentic EBOV. "
[Show abstract][Hide abstract] ABSTRACT: Here, we used a biologically contained Ebola virus system to characterize the spatio-temporal distribution of Ebola virus proteins and RNA during virus replication. We found that viral nucleoprotein (NP), the polymerase cofactor VP35, the major matrix protein VP40, the transcription activator VP30, and the minor matrix protein VP24 were distributed in cytoplasmic inclusions. These inclusions enlarged near the nucleus, became smaller pieces, and subsequently localized near the plasma membrane. GP was distributed in the cytoplasm and transported to the plasma membrane independent of the other viral proteins. We also found that viral RNA synthesis occurred within the inclusions. Newly synthesized negative-sense RNA was distributed inside the inclusions, whereas positive-sense RNA was distributed both inside and outside. These findings provide useful insights into Ebola virus replication.
Full-text · Article · Feb 2013 · Scientific Reports
Steven W Paugh, David R Coss, Ju Bao, Lucas T Laudermilk, Christy R Grace, Antonio M Ferreira, M Brett Waddell, Granger Ridout, Deanna Naeve, Michael Leuze, Philip F LoCascio, John C Panetta, Mark R Wilkinson, Ching-Hon Pui, Clayton W Naeve, Edward C Uberbacher, Erik J Bonten, William E Evans