DOI: 10.5061/dryad.gtht76hsr
title: Data from: The herpes simplex virus pUL16 and pUL21 proteins prevent capsids from docking at nuclear pore complexes

description: After entry into cells, herpes simplex virus (HSV) nucleocapsids dock at
 nuclear pore complexes (NPCs) through which viral genomes are released
 into the nucleoplasm where viral gene expression, genome replication, and
 early steps in virion assembly take place. After their assembly,
 nucleocapsids are translocated to the cytoplasm for final virion
 maturation. Nascent cytoplasmic nucleocapsids are prevented from binding
 to NPCs and delivering their genomes to the nucleus from which they
 emerged, but how this is accomplished is not understood. Here we report
 that HSV pUL16 and pUL21 deletion mutants accumulate empty capsids at the
 cytoplasmic face of NPCs late in infection. Additionally, prior expression
 of pUL16 and pUL21 prevented incoming nucleocapsids from docking at NPCs,
 delivering their genomes to the nucleus and initiating viral gene
 expression. Both pUL16 and pUL21 localized to the nuclear envelope,
 placing them in an appropriate location to interfere with nucleocapsid/NPC
 interactions.
description: # Research data Confocal and TEM images associated with The Herpes Simplex
 Virus pUL16 and pUL21 Proteins Prevent Capsids from Docking at Nuclear
 Pore Complexes ## Description of the data and file structure Folders are
 named with corresponding figure numbers. File types include .oib, .jpg and
 .tif. All can be opened with NIH Image/Fiji. Figure 2D: The localization
 of HSV-2 and HSV-1 capsids in the absence of pUL16 and/or pUL21. The
 confocal images of Vero cells in this folder were used for quantification
 of the percentage of HSV-1 infected cells with capsids accumulated at the
 nuclear envelope. Three biological replicates with n = 198–258 infected
 cells examined per biological replicate. Subfolder titles correspond to
 the different virus strains analyzed. Figure 3E: The colocalization of
 HSV-2 capsids with NPCs at late times in infection. The electron
 micrographs of Vero cells in this folder were used for quantification of
 the number of A-capsids docked at NPCs per nuclear section. n = 10–26
 nuclear sections examined per condition. Subfolder titles correspond to
 the different virus strains analyzed. Figures 4B and 4C: Examination of
 super-infection inhibition in HSV-2 WT, Δ16 and Δ21 infected cells. The
 confocal images of Vero cells in this folder were used for quantification
 of cytoplasmic mCh-VP26 capsids within mock, WT, Δ16 and Δ21 infected
 cells. n = 10 z-projections of infected cells per condition. Additionally,
 these images were used for the quantification of the percentage of
 mCh-VP26 capsids within the nuclear periphery of mock, WT, Δ16 and Δ21
 infected cells. n = 10 z-projections of infected cells per condition.
 Capsids at the nuclear periphery were defined as those abutting the
 Hoechst 33342 signal. Subfolder titles correspond to the different virus
 strains analyzed. Figure 4E: Examination of super-infection inhibition in
 HSV-2 WT, Δ16 and Δ21 infected cells. The confocal images of Vero cells in
 this folder were used for quantification of nuclear WT EdC labelled viral
 genomes within WT, Δ16 and Δ21 infected cells. n = 177–470 infected cells
 examined for fluorescent EdC puncta per biological replicate. Subfolder
 titles correspond to the different virus strains analyzed. Figure 5B: The
 effect of prior ectopic HSV-2 pUL16 and pUL21 expression on HSV-2 and PRV
 capsid recruitment to nuclei and subsequent virus gene expression. The
 confocal images of HeLa cells in this folder were used for measuring the
 average distance of HSV-2 capsids from the nuclear periphery in HeLa cells
 that had been transfected with EGFP alone (control) or in combination with
 HSV-2 pUL16 and/or pUL21 expression plasmids and infected for 2h with
 HSV-2 186 mCh-VP26. n = 43–55 capsids were examined in each transfection
 condition. Subfolder titles correspond to the different expression
 plasmids, or combinations of plasmids, that were transfected into cells
 prior to infection. Within each subfolder are .oif files. These .oif files
 are Olympus Image File descriptors that store metadata for
 multidimensional z-series microscopy images. Each .oif file is accompanied
 by a corresponding oif.files/ folder containing associated data files such
 as .tif images, .roi regions of interest, .pty acquisition settings, .lut
 lookup tables for visualization, and .bmp preview images. Figure 5C: The
 effect of prior ectopic HSV-2 pUL16 and pUL21 expression on HSV-2 and PRV
 capsid recruitment to nuclei and subsequent virus gene expression. he
 confocal images of PK15 cells in this folder were used for measuring the
 average distance of PRV capsids from the nuclear periphery in PK15 cells
 that had been transfected with EGFP alone (control) or in combination with
 HSV-2 pUL16 and/or pUL21 expression plasmids and infected for 2h with
 PRV765. n = 46–60 capsids were examined in each transfection condition.
 Subfolder titles correspond to the different virus strains analyzed.
 Figure 6B: The colocalization of pUL16-EGFP and pUL21-EGFP with HSV-2
 mCh-VP26 capsids. The confocal images of HeLa cells in this folder were
 used for quantifying the colocalization between EGFP signal and mCh-VP26
 capsid signals compared to the average EGFP background signal taken from
 two points 0.5μm from the analyzed capsid. n = 40 capsids per transfection
 condition were examined. No significant differences were seen between any
 of the transfection conditions when comparing the EGFP signal associated
 with mCh-VP26 capsids to the EGFP control. Subfolder titles correspond to
 the different virus strains analyzed. Figure 7E: The colocalization of HSV
 capsids with NPCs in cells treated with nocodazole. The confocal images of
 Vero cells in this folder were used for quantification of nuclei with
 capsids colocalizing with NPCs in HSV-2 infected cells treated with DMSO
 (vehicle control) or nocodazole at 12 hpi and fixed at 18 hpi. Three
 biological replicates with n = 100–140 infected cells examined per
 biological replicate. Subfolder titles correspond to the different virus
 strains analyzed in the presence and absence of nocodazole. ##
 Sharing/Access information Files can be viewed with NIH Image/Fiji. The
 Bio-Formats plugin is required to see the .oib files in Fiji. ##
 Code/Software N/A