Involvement of actin-containing microfilaments in HSV-induced cytopathology and the influence of inhibitors of glycosylation

Heeg, U.; Dienes, Hans‐Peter; Müller, Sabine; Falke, D.

doi:10.1007/bf01314285

Cited by 33 publications

(27 citation statements)

References 29 publications

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“…4A). Actin stress fibers are also largely depolymerized during infection (16,41,59,63). When infected Vero cells were stained for actin with Texas Red-labeled phalloidin, few stress fibers were visible but the actin cortex (the unbundled, highly branched actin lining the cell membrane) could still be seen in most cases (Fig.…”

Section: Resultsmentioning

confidence: 99%

Replication of Herpes Simplex Virus: Egress of Progeny Virus at Specialized Cell Membrane Sites

Mingo

Han

Newcomb

et al. 2012

J Virol

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bIn the final stages of the herpes simplex virus 1 (HSV-1) life cycle, a viral nucleocapsid buds into a vesicle of trans-Golgi network (TGN)/endosome origin, acquiring an envelope and an outer vesicular membrane. The virus-containing vesicle then traffics to the plasma membrane where it fuses, exposing a mature virion. Although the process of directed egress has been studied in polarized epithelial cell lines, less work has been done in nonpolarized cell types. In this report, we describe a study of HSV-1 egress as it occurs in nonpolarized cells. The examination of infected Vero cells by electron, confocal, and total internal reflection fluorescence (TIRF) microscopy revealed that HSV-1 was released at specific pocket-like areas of the plasma membrane that were found along the substrate-adherent surface and cell-cell-adherent contacts. Both the membrane composition and cytoskeletal structure of egress sites were found to be modified by infection. The plasma membrane at virion release sites was heavily enriched in viral glycoproteins. Small glycoprotein patches formed early in infection, and virus became associated with these areas as they expanded. Glycoprotein-rich areas formed independently from virion trafficking as confirmed by the use of a UL25 mutant with a defect in capsid nuclear egress. The depolymerization of the cytoskeleton indicated that microtubules were important for the trafficking of virions and glycoproteins to release sites. In addition, the actin cytoskeleton was found to be necessary for maintaining the integrity of egress sites. When actin was depolymerized, the glycoprotein concentrations dispersed across the membrane, as did the surface-associated virus. Lastly, viral glycoprotein E appeared to function in a different manner in nonpolarized cells compared to previous studies of egress in polarized epithelial cells; the total amount of virus released at egress sites was slightly increased in infected Vero cells when gE was absent. However, gE was important for egress site formation, as Vero cells infected with gE deletion mutants formed glycoprotein patches that were significantly reduced in size. The results of this study are interpreted to indicate that the egress of HSV-1 in Vero cells is directed to virally induced, specialized egress sites that form along specific areas of the cell membrane.

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Section: Resultsmentioning

confidence: 99%

Replication of Herpes Simplex Virus: Egress of Progeny Virus at Specialized Cell Membrane Sites

Mingo

Han

Newcomb

et al. 2012

J Virol

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“…CV-1 cells were grown either on coverslips or in flasks at a density of 3 x 104 cells/cm 2 (Bachmann et al, 1989). The~, were infected with HSV-1 strains HFEM, Lenette or 17 syn ÷ or with temperaturesensitive (ts) mutants at high multiplicity (10 p.f.u,/cell) (Heeg et at., 1986;Metzger et al, 1988). The ts mutants were defective in either the immediate early (IE) protein (M~ 175K) (tsK) or in DNA synthesis (tsS) (Metzger et al, 1988).…”

Section: Methodsmentioning

confidence: 99%

Intracellular Distribution of the La Antigen in CV-1 Cells after Herpes Simplex Virus Type 1 Infection Compared with the Localization of U Small Nuclear Ribonucleoprotein Particles

Bachmann

Falke

Schröder

et al. 1989

Journal of General Virology

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SUMMARYThe La antigen is known to associate, at least transiently, with a series of small nuclear and cytoplasmic ribonucleoprotein particles (snRNPs and scRNPs), e.g. U1 and U6 snRNPs. In CV-1 cells a monoclonal antibody (MAb), directed against the La protein (LalB5), immunostained intranuclear speckles. These speckles were found to co-localize with speckles that were stained by MAbs directed against either all U snRNPs or only against U1 snRNPs. Two h after infection of CV-1 cells with herpes simplex virus type 1 (HSV-1) (strain HFEM) the staining of nuclear speckles with the anti-La MAb disappeared and the La protein was found quantitatively in the cytoplasm. In contrast nuclear speckles remained stained with the MAbs against the U snRNPs. Similar results were obtained using HSV-1 strains Lenette or 17 syn ÷ or temperature-sensitive (ts) mutants defective either in DNA synthesis (tsS) or in the immediate early protein (Mr 175 K) (tsK). Later in infection the La protein returned to the nucleus. Six h after infection most of the nuclear La protein was found to localize within patchy regions. These areas seem to be related to heterogeneous nuclear RNA transcription and/or processing sites, but not to DNA replication sites.

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“…This cytopathology is characterized by (i) rounding up of cells caused by cytoskeletal destabilizations and loss of matrix binding proteins, (ii) nucleolar alterations and chromatin margination or damage, and (iii) an overall decrease in cellular macromolecular synthesis (5,25,26,55,57,58). Although cytolysis due to viral replication is generally believed to occur through a necrotic route, recent studies have indicated that one aspect of HSV-1-induced CPE is programmed cell death, or apoptosis (1,3,19,32,35).…”

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Herpes Simplex Virus Type 1 Immediate-Early Gene Expression Is Required for the Induction of Apoptosis in Human Epithelial HEp-2 Cells

Sanfilippo

Chirimuuta

Blaho

2004

J Virol

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Wild-type herpes simplex virus type 1 (HSV-1) induces apoptosis in human epithelialHerpes simplex virus type 1 (HSV-1) is an important pathogen that causes a variety of clinical manifestations in humans. It has the ability to remain latent in host neurons for life and reactivate to cause lesions at or near the site of initial infection. Reactivation from the latent state results in productive infection that ultimately leads to the lytic destruction of distal epithelial cells. During the lytic cycle of the virus in cultured cells, regulation of HSV-1 replication occurs mainly at the transcriptional level and involves the coordination of three phases of gene expression (reviewed in reference 57). Viral genes are expressed in a tightly regulated, ordered cascade (27, 28), which begins with the production of the immediate-early (IE), or ␣, genes. The resulting ␣/IE proteins, which include infected cell proteins ICP0, ICP4, ICP22, and ICP27, are responsible for regulating viral gene expression during subsequent phases of the replication cycle (57). Transcription of the ␣ genes occurs in the absence of de novo viral protein synthesis (8) and is highly stimulated by the ␣ trans-inducing activity of virion VP16 (13, 50, 52). The early (␤) gene products, for example, the viral thymidine kinase (TK), are synthesized next and are principally involved in viral DNA synthesis (reviewed in reference 12). The last set of genes expressed are the late (␥) genes, which encode proteins involved in virion structure and assembly, such as VP16, the virion host shutoff (vhs) protein, VP22, and glycoprotein C (gC) (reviewed in reference 17).Productive HSV-1 replication induces major biochemical changes in infected cells that are collectively referred to as cytopathic effect (CPE). This cytopathology is characterized by (i) rounding up of cells caused by cytoskeletal destabilizations and loss of matrix binding proteins, (ii) nucleolar alterations and chromatin margination or damage, and (iii) an overall decrease in cellular macromolecular synthesis (5,25,26,55,57,58). Although cytolysis due to viral replication is generally believed to occur through a necrotic route, recent studies have indicated that one aspect of HSV-1-induced CPE is programmed cell death, or apoptosis (1,3,19,32,35). Apoptosis is a highly regulated process of cell suicide that is induced by death signals received from either a cell surface death receptor or the mitochondria (22,23,47,67). The process of apoptosis involves a family of aspartate-specific cysteinyl proteases, or caspases, which are activated by proteolytic cleavage. Regardless of the source from which a death signal originates, downstream "executioner" caspases, such as caspase-3, are common to both pathways (22,59,63,68). This subclass of caspases is responsible for the processing of various cytoplasmic and nuclear substrates, such as the DNA repair enzyme poly(ADPribose) polymerase (PARP), a 116-kDa protein which generates an 85-kDa product upon processing (59). The cascade of events culminating in cas...

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Involvement of actin-containing microfilaments in HSV-induced cytopathology and the influence of inhibitors of glycosylation

Cited by 33 publications

References 29 publications

Replication of Herpes Simplex Virus: Egress of Progeny Virus at Specialized Cell Membrane Sites

Replication of Herpes Simplex Virus: Egress of Progeny Virus at Specialized Cell Membrane Sites

Intracellular Distribution of the La Antigen in CV-1 Cells after Herpes Simplex Virus Type 1 Infection Compared with the Localization of U Small Nuclear Ribonucleoprotein Particles

Herpes Simplex Virus Type 1 Immediate-Early Gene Expression Is Required for the Induction of Apoptosis in Human Epithelial HEp-2 Cells

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