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Analysis of rescue efficiency, replication, and matrix localization of recombinant Sendai virus bearing matrix nuclear export mutants.

By Mickey Pentecost (237685), Ajay A. Vashisht (128117), Talia Lester (703968), Tim Voros (703969), Shannon M. Beaty (703970), Arnold Park (237682), Yao E. Wang (237680), Tatyana E Yun (703971), Alexander N. Freiberg (95391), James A. Wohlschlegel (212752) and Benhur Lee (78126)

Abstract

<p><b>(A)</b> Comparison of virus-like particle (VLP) budding between 3X-Flag-tagged NiV-M, HeV-M, MuV-M, and SeV-M. Anti-Flag immunoblots of cell lysate and purified VLPs from HEK 293T cells 24 h post transfection with the indicated constructs. Normalized budding index was calculated from immunoblot integrated intensities. <b>(B)</b> Quantification of rescue events (GFP+ cells) from three independent rescues of rSeV-eGFP containing WT, L102 L103 mutant, or K254R mutant SeV-M at day 2 post rescue in HEK 293T cells. <b>(C)</b> Quantification of viral titers from the rescues of rSeV-eGFP containing WT, L102 L103 mutant, or K254R mutant SeV-M at day 6 post rescue in HEK 293T cells. <b>(D)</b> Extended Focus (maximum intensity projection) view of 3D confocal micrographs of HEK 293T cells at day 6 post rescue of rSeV-eGFP containing WT, L102 L103 mutant, or K254R mutant SeV-M. Cells were counterstained with anti-SeV-M antibodies, red. <b>(E)</b> Extended Focus (maximum intensity projection) view of 3D confocal micrographs of HEK 293T cells at day 6 post rescue of rSeV-eGFP containing WT, L102 L103 mutant, or K254R mutant SeV-M. Cells were counterstained with anti-SeV-F antibodies, red. <b>(F)</b> XYZ Planes View of 3D confocal micrographs of HEK 293T cells at day 6 post rescue of rSeV-eGFP containing WT, L102 L103 mutant, or K254R mutant SeV-M. Cells were counterstained with DAPI to visualize nuclei, blue, anti-SeV-M antibodies, red, and anti-Fibrillarin antibodies to visualize nucleoli, grayscale.</p

Topics: Biological Sciences, Nipah virus M, Paramyxovirinae Matrix Proteins, M proteins hints, shotgun proteomics experiments, Paramyxovirinae M proteins, K 258 residue, nes, Recombinant Mumps virus, 3 D confocal microscopy, M proteins, paramyxovirus M proteins, Recombinant Sendai virus, trafficking
Year: 2015
DOI identifier: 10.1371/journal.ppat.1004739.g007
OAI identifier: oai:figshare.com:article/1339557
Provided by: FigShare
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