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Comparative Study
. 2001 Apr;75(8):3675-84.
doi: 10.1128/JVI.75.8.3675-3684.2001.

Egress of alphaherpesviruses: comparative ultrastructural study

Affiliations
Comparative Study

Egress of alphaherpesviruses: comparative ultrastructural study

H Granzow et al. J Virol. 2001 Apr.

Abstract

Egress of four important alphaherpesviruses, equine herpesvirus 1 (EHV-1), herpes simplex virus type 1 (HSV-1), infectious laryngotracheitis virus (ILTV), and pseudorabies virus (PrV), was investigated by electron microscopy of infected cell lines of different origins. In all virus-cell systems analyzed, similar observations were made concerning the different stages of virion morphogenesis. After intranuclear assembly, nucleocapsids bud at the inner leaflet of the nuclear membrane, resulting in enveloped particles in the perinuclear space that contain a sharply bordered rim of tegument and a smooth envelope surface. Egress from the perinuclear cisterna primarily occurs by fusion of the primary envelope with the outer leaflet of the nuclear membrane, which has been visualized for HSV-1 and EHV-1 for the first time. The resulting intracytoplasmic naked nucleocapsids are enveloped at membranes of the trans-Golgi network (TGN), as shown by immunogold labeling with a TGN-specific antiserum. Virions containing their final envelope differ in morphology from particles within the perinuclear cisterna by visible surface projections and a diffuse tegument. Particularly striking was the addition of a large amount of tegument material to ILTV capsids in the cytoplasm. Extracellular virions were morphologically identical to virions within Golgi-derived vesicles, but distinct from virions in the perinuclear space. Studies with gB- and gH-deleted PrV mutants indicated that these two glycoproteins, which are essential for virus entry and direct cell-to-cell spread, are dispensable for egress. Taken together, our studies indicate that the deenvelopment-reenvelopment process of herpesvirus maturation also occurs in EHV-1, HSV-1, and ILTV and that membrane fusion processes occurring during egress are substantially different from those during entry and direct viral cell-to-cell spread.

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Figures

FIG. 1
FIG. 1
Exit of nucleocapsids from the nucleus. Enveloped EHV-1 (A), HSV-1 (B), ILTV (C), and PrV (D) particles in the perinuclear cisterna are shown. Release of EHV-1 (E), HSV-1 (F), and PrV (H) from the nuclear cisterna results from direct fusion. Simultaneous observation of primary envelopment (arrowhead) and deenvelopment (arrow) of HSV-1 particles is demonstrated in panel G. Capsidless HSV-1 particles (arrowhead) in the perinuclear cisterna (I) and fused with the outer lamella of the nuclear membrane (J [arrowhead]) were also observed. The bars represent 150 nm in panels A to F and H and 200 nm in panels G and I. N designates the nucleus.
FIG. 2
FIG. 2
Secondary envelopment and virus egress. Envelopment of EHV-1 (A), HSV-1 (B), ILTV (C), and PrV (D) at membranes of the trans-Golgi area. Particles of EHV-1 (E), HSV-1 (F), ILTV (G), and PrV (H) within Golgi vesicles are shown, as is egress of EHV-1 (I), HSV-1 (J), ILTV (K), and PrV (L) by fusion of vesicles with the plasma membrane. The bars represent 150 nm in panels A to K and 250 nm in panel L.
FIG. 3
FIG. 3
Overview of Vero cells infected with HSV-1 (A) and LMH cells infected with ILTV (B). The inset shows the formation of an ILTV L-particle. The bars represent 1.5 μm in panels A and B and 150 nm in the panel B inset.
FIG. 4
FIG. 4
Immunolabeling of TGN38 protein. Ultrathin sections of Lowicryl-embedded PrV-infected porcine kidney cells were processed as described in Materials and Methods and incubated with two different dilutions (A, 1:100; B, 1:1,000) of anti-TGN38 serum (44) followed by 10-nm-diameter gold particle-tagged secondary antibodies. The bars represent 250 nm.
FIG. 5
FIG. 5
Morphology of virus particles. Virions in the perinuclear cisterna after primary envelopment: EHV-1 (A), HSV-1 (B), ILTV (C), and PrV (D); virions in Golgi vesicles after secondary envelopment: EHV-1 (E), HSV-1 (F), ILTV (G), and PrV (H); extracellular virions after virus egress: EHV-1 (I), HSV-1 (J), ILTV (K), and PrV (L). Bars, 150 nm.
FIG. 6
FIG. 6
Ultrastructure of cells infected with gB or gH deletion mutants of PrV. Noncomplementing RK13 cells were infected with PrV-gB (A) or PrV-gH (B) at an MOI of 1 and analyzed 16 h after infection. All stages of virus maturation, including numerous extracellular virus particles, can be observed. The bars represent 1.5 μm in panels A and B and 500 nm in the insets.

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References

    1. Babic N, Klupp B G, Makoschey B, Karger A, Flamand A, Mettenleiter T C. Glycoprotein gH of pseudorabies virus is essential for penetration and propagation in cell culture and in the nervous system of mice. J Gen Virol. 1996;77:2277–2285. - PubMed
    1. Brack A R, Dijkstra J M, Granzow H, Klupp B G, Mettenleiter T C. Inhibition of virion maturation by simultaneous deletion of glycoproteins E, I, and M of pseudorabies virus. J Virol. 1999;73:5364–5372. - PMC - PubMed
    1. Browne H, Bell S, Minson T, Wilson D W. An endoplasmic reticulum-retained herpes simplex virus glycoprotein H is absent from secreted virions: evidence for reenvelopment during egress. J Virol. 1996;70:4311–4316. - PMC - PubMed
    1. Brunetti C R, Dingwell K S, Wale C, Graham F L, Johnson D C. Herpes simplex virus gD and virions accumulate in endosomes by mannose 6-phosphate-dependent and -independent mechanisms. J Virol. 1998;72:3330–3339. - PMC - PubMed
    1. Campadelli-Fiume G, Arsenakis M, Farabegoli F, Roizman B. Entry of herpes simplex virus in BJ cells that constitutively express viral glycoprotein D is by endocytosis and results in degradation of the virus. J Virol. 1988;62:159–167. - PMC - PubMed

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