Characterization of a Herpes Simplex Virus type 1 Deletion Variant (1703) Which Under-produces Vmw63 During Immediate Early Conditions of Infection

Mc, Sinclair; McLauchlan, John; Marsden, Howard S.; Sm, Brown

doi:10.1099/0022-1317-75-5-1083

Cited by 6 publications

(3 citation statements)

References 25 publications

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“…K. Artzt (University of Texas, Austin, Texas), anti-QKI-Pan clone N147/6 (UC Davis/NIH NeuroMab Facility), anti-US11 (27), anti-KDEL, anti-ICP4, anti-ICP27, and anti-␤-actin (Abcam, Cambridge, MA), anti-histone H4 (Millipore, Billerica, MA), anti-p27 Kip1 , anti-p57 Kip2 (Cell Signaling, Danvers, MA), antinectin-1, anti-HDAC-1, anti-HNF␣, and anti-ICP0 (Santa Cruz Biotechnology, Santa Cruz, CA). Anti-UL42 was kindly provided by Dr. H. Marsden (28). Electrophoresis reagents were purchased from GE Healthcare and trypsin from Promega (Madison, WI).…”

Section: Methodsmentioning

confidence: 99%

HSV-1 Cgal+ Infection Promotes Quaking RNA Binding Protein Production and Induces Nuclear-Cytoplasmic Shuttling of Quaking I-5 Isoform in Human Hepatoma Cells

Sanchez-Quiles

Mora

Segura

et al. 2011

Molecular & Cellular Proteomics

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Herpesvirus type 1 (HSV-1) based oncolytic vectors arise as a promising therapeutic alternative for neoplastic diseases including hepatocellular carcinoma. However, the mechanisms mediating the host cell response to such treatments are not completely known. It is well established that HSV-1 infection induces functional and structural alterations in the nucleus of the host cell. In the present work, we have used gel-based and shotgun proteomic strategies to elucidate the signaling pathways impaired in the nucleus of human hepatoma cells (Huh7) upon HSV-1 Cgal(+) infection. Both approaches allowed the identification of differential proteins suggesting impairment of cell functions involved in many aspects of host-virus interaction such as transcription regulation, mRNA processing, and mRNA splicing. Based on our proteomic data and additional functional studies, cellular protein quaking content (QKI) increases 4 hours postinfection (hpi), when viral immediate-early genes such as ICP4 and ICP27 could be also detected. Depletion of QKI expression by small interfering RNA results in reduction of viral immediate-early protein levels, subsequent decrease in early and late viral protein content, and a reduction in the viral yield indicating that QKI directly interferes with viral replication. In particular, HSV-1 Cgal(+) induces a transient increase in quaking I-5 isoform (QKI-5) levels, in parallel with an enhancement of p27(Kip1) protein content. Moreover, immunofluorescence microscopy showed an early nuclear redistribution of QKI-5, shuttling from the nucleus to the cytosol and colocalizing with nectin-1 in cell to cell contact regions at 16-24 hpi. This evidence sheds new light on mechanisms mediating hepatoma cell response to HSV-1 vectors highlighting QKI as a central molecular mediator.

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Section: Methodsmentioning

confidence: 99%

HSV-1 Cgal+ Infection Promotes Quaking RNA Binding Protein Production and Induces Nuclear-Cytoplasmic Shuttling of Quaking I-5 Isoform in Human Hepatoma Cells

Sanchez-Quiles

Mora

Segura

et al. 2011

Molecular & Cellular Proteomics

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“…Viral proteins (10 µg aliquots) were analyzed by Western blots using a 50× dilution of either a rabbit polyclonal anti‐ ICP27 antibody, or anti‐Us11 antibody (28), or a mouse monoclonal anti‐ UL42 antibody. Anti‐ ICP27 and anti‐ UL42 antibodies were kindly provided by Dr. Marsden (antibodies 42 and Z1F11, respectively) (29, 30). Proteins were revealed by ECL (Amersham Biosciences) using an anti‐rabbit or an anti‐mouse peroxydase‐conjugate (Sigma) diluted 1:10000.…”

Section: Methodsmentioning

confidence: 99%

S‐adenosyl methionine decarboxylase activity is required for the outcome of herpes simplex virus type 1 infection and represents a new potential therapeutic target

et al. 2005

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All the available antiherpetic drugs are directed against viral proteins. Their extensive clinical use has led to the emergence of resistant viral strains. There is a need for the treatment of herpes infections due to resistant strains, especially for immunocompromised patients. To design new kinds of drugs, we have developed a strategy to identify cellular targets. Herpes simplex virus type 1 (HSV-1) infection is concomitant to a repression of most host protein synthesis. However, some cellular proteins continue to be efficiently synthesized. We speculated that some of them could determine the outcome of infection. Since two polyamines, spermidine and spermine, are components of the HSV-1 virions, we investigated whether enzymes involved in their synthesis could be required for viral infection. We show that inhibition of S-adenosyl methionine decarboxylase, a key enzyme of the polyamine metabolic pathway, prevents HSV-1 infection. Inhibition of polyamine synthesis prevents infection of culture cells with HSV-1 laboratory strains as well as clinical isolates that are resistant to the conventional antiviral drugs acyclovir and foscarnet. Our data provide the opportunity to develop molecules with a novel mechanism of action for the treatment of herpes infection.

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“…The membranes were incubated with a 500ϫ dilution of either a rabbit polyclonal antinucleolin antibody (19); antifibrillarin (N-15; Santa Cruz), anti-ICP27, or anti-US11 antibodies (14); or mouse monoclonal anti-B23 (H-106; Sigma), anti-␤-actin (AC-15; Sigma), antihistone H3 (Abcam), or anti-UL42 antibodies. Anti-ICP27 and anti-UL42 antibodies were kindly provided by H. Marsden (antibodies 42 and Z1F11, respectively) (53,58). Proteins were revealed by chemiluminescence (ECL from Amersham Biosciences) using an anti-rabbit or an anti-mouse peroxidase-conjugated antibody (Sigma) or an anti-goat peroxidase-conjugated antibody (Santa Cruz), diluted 1:10,000.…”

Section: Methodsmentioning

confidence: 99%

Nucleolin Is Required for an Efficient Herpes Simplex Virus Type 1 Infection

Callé

Ugrinova

Epstein

et al. 2008

J Virol

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Productive infection by herpes simplex virus type 1 (HSV-1), which occurs in the host cell nucleus, is accompanied by dramatic modifications of the nuclear architecture, including profound alterations of nucleolar morphology. Here, we show that the three most abundant nucleolar proteins-nucleolin, B23, and fibrillarin-are redistributed out of the nucleoli as a consequence of HSV-1 infection. We show that the amount of nucleolin increases progressively during the course of infection. We demonstrate for the first time that a nucleolar protein, i.e., nucleolin, colocalizes with ICP8 in the viral replication compartments, at the time when viral replication is effective, suggesting an involvement of nucleolin in the HSV-1 DNA replication process. At later times of infection, a granular form of nucleolin localizes to the cytoplasm, in structures that display the characteristic features of aggresomes, indicating that this form of nucleolin is very probably destined for degradation. The delocalization of nucleolin from the nucleoli requires the viral ICP4 protein or a factor(s) whose expression involves ICP4. Using small interfering RNA technology, we show that viral replication requires a high level of nucleolin expression, demonstrating for the first time a direct role for a nucleolar protein in herpes simplex virus biology.Herpes simplex virus type 1 (HSV-1) is a human herpesvirus consisting of an outer envelope, a tegument, a capsid, and a linear double-stranded DNA. Productive infection consists of a highly ordered program of viral gene expression, DNA replication, and virion assembly that leads to the formation of infectious viral progeny and cell death (48). The viral DNA contains at least 80 genes whose expression is sequentially and temporally regulated by complex regulatory mechanisms. Viral genes can be divided into immediate-early, early, and late genes, according to their kinetics of expression. Proteins encoded by immediate-early genes are involved in the regulation of the synthesis of early and late proteins. Proteins encoded by early genes participate in viral DNA replication, and proteins encoded by late genes are mainly the structural components of the viral particles.Viral transcription, DNA replication, assembly of new capsids, and packaging of HSV-1 DNA occur in the host cell nucleus. As a consequence, HSV-1-infected cells undergo a variety of changes including dramatic modifications of the nuclear architecture. The formation of viral replication compartments (VRC), which are the sites of replication, transcription, and encapsidation of HSV-1 genomes, is accompanied by the marginalization of chromatin and the disruption of the nuclear lamina and of PML bodies, as well as by a profound modification of nucleolar morphology (2,15,33,39). Soon after infection, nucleoli increase in size, localize close to the nuclear membrane, and finally become fragmented in small pieces (39,49). In addition, several viral proteins, including ICP0, ICP4, ICP27, US11, and gamma 34.5, localize at least transiently to nucl...

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Characterization of a Herpes Simplex Virus type 1 Deletion Variant (1703) Which Under-produces Vmw63 During Immediate Early Conditions of Infection

Cited by 6 publications

References 25 publications

HSV-1 Cgal+ Infection Promotes Quaking RNA Binding Protein Production and Induces Nuclear-Cytoplasmic Shuttling of Quaking I-5 Isoform in Human Hepatoma Cells

HSV-1 Cgal+ Infection Promotes Quaking RNA Binding Protein Production and Induces Nuclear-Cytoplasmic Shuttling of Quaking I-5 Isoform in Human Hepatoma Cells

S‐adenosyl methionine decarboxylase activity is required for the outcome of herpes simplex virus type 1 infection and represents a new potential therapeutic target

Nucleolin Is Required for an Efficient Herpes Simplex Virus Type 1 Infection

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