A block in glycoprotein processing correlates with small plaque morphology and virion targetting to cell-cell junctions for an oral and an anal strain of herpes simplex virus type-1.
*Protein Processing; Anal Canal/virology; Animals; Cercopithecus aethiops; Electron; Fluorescent Antibody Technique; Genetic Complementation Test; Herpesvirus 1; Human/genetics/isolation & purification/*physiology; Humans; Indirect; Intercellular Junctions/physiology/*virology; Kinetics; Microscopy; Mouth/virology; Post-Translational; Species Specificity; Vero Cells; Viral Envelope Proteins/*biosynthesis/metabolism; Viral Plaque Assay; Virion/*physiology
The characteristics of two clinical isolates of HSV-1 obtained from an oral (424) and an anal (490) lesion were compared with the highly passaged KOS strain. In contrast to KOS, the clinical isolates produced small plaques, were more cell-associated and the predominant viral glycoprotein species for gC and gD in infected cell lysates was the precursor, high mannose glycoform. Total virus production in Vero cells was equivalent for the three virus strains in one-step growths. Pulse-chase studies of glycoprotein C processing showed a reduction in rate at 7.5 h post infection and a significant block in processing at 10.5 h post infection for 424 and 490 but not KOS. Similar results were obtained for gD. The significant reduction in glycoprotein processing for 424 and 490 suggests a block in transport of viral glycoproteins or virions to and through the Golgi apparatus. Extracellular virions and the cell surface, prior to cell lysis, contained the processed gC glycoform suggesting a competent cellular glycan processing system. Upon co-infection of 424 or 490 with KOS or a gC- KOS strain, gC was processed to levels equivalent to KOS indicating that 424 and 490 are not inhibitory but that an activity(s) encoded by KOS facilitates maturation of gC from 424 and 490. Unlike KOS infected Vero cells, virion-containing vacuoles were observed in the cytoplasm at 12 h p.i. and extracellular virions were concentrated at cell-cell junctions of 424 or 490 infected cells but not in the perinuclear region. These results suggest that intracellular transport of viral glycoproteins and virions in 424 and 490 infected cells is different from KOS infected cells. The reduced level of viral glycoprotein maturation, virus release, cell surface presence and presence of virions at cell-cell junctions are consistent with small plaque production in tissue culture cells.
Dick J W; Rosenthal K S
Archives of virology
1995
1995
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/bf01323238" target="_blank" rel="noreferrer noopener">10.1007/bf01323238</a>
Mice transgenic for simian immunodeficiency virus nef are immunologically compromised.
*Genes; *Herpesvirus 1; *Immunocompromised Host; 3T3 Cells; Aging; Animals; Antibodies; Antibody Formation; Cellular; Cercopithecus aethiops; Female; Herpes Simplex/*immunology; HIV/pathogenicity; Human; Humans; Immunity; Immunoglobulin G/blood; Kinetics; Male; Mice; nef; Simian Immunodeficiency Virus/*genetics/pathogenicity; Transgenic; Vero Cells; Viral/biosynthesis/blood
An intact nef gene is essential for rapid development of immunodeficiency in human immunodeficiency virus and simian immunodeficiency virus infections. To assess the role of nef in the immune response, mice transgenic for SIV nef were constructed and the humoral and cellular immune response to herpes simplex virus type-1 (HSV-1), measured. Mice transgenic for SIVmac239 nef exhibited a significantly increased mortality rate when challenged with HSV-1 and also showed unusual antibody kinetics in response to viral challenge. During a 32-week period following exposure to HSV, it was noted that IgG subclass titers continued to rise in the nef+ animals, while titers of nef- animals decreased. Additionally, following secondary challenge with HSV, nef- mice had a significantly greater rise in HSV-neutralizing antibody titers than nef+ mice. A decreased proliferative response to the T cell mitogen, PHA, was noted in the nef+ animals. These results suggest that the presence of nef+ is sufficient to induce immune dysfunction.
Larsen N B; Kestler H W; Docherty J J
Journal of biomedical science
1998
1998-08
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/bf02255857" target="_blank" rel="noreferrer noopener">10.1007/bf02255857</a>
HSV-2 ICP34.5 protein modulates herpes simplex virus glycoprotein processing.
*Protein Processing; Animals; Cercopithecus aethiops; Cloning; Gene Expression; Herpesvirus 1; Herpesvirus 2; Human/*genetics; Molecular; Post-Translational; Vero Cells; Viral Envelope Proteins/*metabolism; Viral Proteins/genetics/*metabolism
The ICP34.5 gene from HSV-2 strain 333 was cloned and, when expressed in Vero cells, enhanced the efficiency and extent of glycoprotein processing of glycoprotein C (gC1), a representative viral glycoprotein, during infection with
Chatterjee Somik; Wang Jason W; Cismowski Mary J; Bower John R; Rosenthal Kenneth Steven
Archives of virology
2009
1905-07
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s00705-009-0341-9" target="_blank" rel="noreferrer noopener">10.1007/s00705-009-0341-9</a>
The large protein 'L' of Peste-des-petits-ruminants virus exhibits RNA triphosphatase activity, the first enzyme in mRNA capping pathway.
Acid Anhydride Hydrolases/*metabolism; Animals; Baculoviridae/genetics; Cercopithecus aethiops; Cloning; Conventional mRNA capping; Enzyme Activation; Gene Expression; Genetic Vectors/genetics; Messenger/*genetics/*metabolism; Molecular; Morbillivirus; mRNA capping; Peste-des-petits-ruminants virus L protein; Peste-des-petits-ruminants virus/*physiology; Peste-des-Petits-Ruminants/*virology; PPRV; RNA; RNA Caps/*metabolism; RNA triphosphatase; Vero Cells; Viral Proteins/*metabolism
Peste-des-petits-ruminants is a highly contagious and fatal disease of goats and sheep caused by non-segmented, negative strand RNA virus belonging to the Morbillivirus genus-Peste-des-petits-ruminants virus (PPRV) which is evolutionarily closely related to Rinderpest virus (RPV). The large protein 'L' of the members of this genus is a multifunctional catalytic protein, which transcribes and replicates the viral genomic RNA as well as possesses mRNA capping, methylation and polyadenylation activities; however, the detailed mechanism of mRNA capping by PPRV L protein has not been studied. We have found earlier that the L protein of RPV has RNA triphosphatase (RTPase), guanylyltransferase (GTase) and methyltransferase activities, and unlike vesicular stomatitis virus (VSV), follows the conventional pathway of mRNA capping. In the present work, using a 5'-end labelled viral RNA as substrate, we demonstrate that PPRV L protein has RTPase activity when present in the ribonucleoprotein complex of purified virus as well as recombinant L-P complex expressed in insect cells. Further, a minimal domain in the C-terminal region (aa1640-1840) of the L protein has been expressed in E. coli and shown to exhibit RTPase activity. The RTPase activity of PPRV L protein is metal-dependent and functions with a divalent cation, either magnesium or manganese. In addition, RTPase associated nucleotide triphosphatase activity (NTPase) of PPRV L protein is also demonstrated. This work provides the first detailed study of RTPase activity and identifies the RTPase domain of PPRV L protein.
Ansari Mohammad Yunus; Singh Piyush Kumar; Rajagopalan Deepa; Shanmugam Purnima; Bellur Asutosh; Shaila Melkote Subbarao
Virus Genes
2019
2019-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s11262-018-1617-5" target="_blank" rel="noreferrer noopener">10.1007/s11262-018-1617-5</a>
Resveratrol suppresses nuclear factor-kappaB in herpes simplex virus infected cells.
Animals; Antiviral Agents/*pharmacology; Cell Nucleus/chemistry; Cercopithecus aethiops; Cytoplasm/chemistry; DNA; Electrophoretic Mobility Shift Assay; Fluorescence; Herpesvirus 1; Herpesvirus 2; Human/drug effects/genetics/*growth & development; Human/drug effects/genetics/*physiology; I-kappa B Proteins/metabolism; Messenger/biosynthesis; Microscopy; NF-kappa B/*metabolism; NF-KappaB Inhibitor alpha; Polymerase Chain Reaction; Resveratrol; Reverse Transcriptase Polymerase Chain Reaction; RNA; Stilbenes/*pharmacology; Transcription Factor RelA/metabolism; Vero Cells; Viral/biosynthesis; Virus Replication/*drug effects
Resveratrol inhibits herpes simplex virus (HSV) replication by an unknown mechanism. Previously it was suggested that this inhibition may be mediated through a cellular factor essential for HSV replication [Docherty, J.J., Fu, M.M., Stiffler, B.S., Limperos, R.J., Pokabla, C.M., DeLucia, A.L., 1999. Resveratrol inhibition of herpes simplex virus replication. Antivir. Res. 43,
Faith Seth A; Sweet Thomas J; Bailey Erin; Booth Tristan; Docherty John J
Antiviral research
2006
2006-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.antiviral.2006.06.011" target="_blank" rel="noreferrer noopener">10.1016/j.antiviral.2006.06.011</a>
Resveratrol inhibition of herpes simplex virus replication.
Animals; Antiviral Agents/*pharmacology/toxicity; Cell Cycle/drug effects; Cell Line; Cercopithecus aethiops; Herpesvirus 1; Herpesvirus 2; Human/*drug effects/physiology; Humans; Immediate-Early Proteins/antagonists & inhibitors/biosynthesis; Mice; Resveratrol; Stilbenes/*pharmacology/toxicity; Vero Cells; Virus Latency/drug effects/physiology; Virus Replication/*drug effects/physiology
Resveratrol, a phytoalexin, was found to inhibit herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) replication in a dose-dependent, reversible manner. The observed reduction in virus yield was not caused by the direct inactivation of HSV by resveratrol nor inhibition of virus attachment to the cell. The chemical did, however, target an early event in the virus replication cycle since it was most effective when added within 1 h of cell infection, less effective if addition was delayed until 6 h post-infection and not effective if added 9 h post-infection. Resveratrol was also found to delay the cell cycle at S-G2-M interphase, inhibit reactivation of virus from latently-infected neurons and reduce the amount of ICP-4, a major immediate early viral regulatory protein, that is produced when compared to controls. These results suggest that a critical early event in the viral replication cycle, that has a compensatory cellular counterpart, is being adversely affected.
Docherty J J; Fu M M; Stiffler B S; Limperos R J; Pokabla C M; DeLucia A L
Antiviral research
1999
1999-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/s0166-3542(99)00042-x" target="_blank" rel="noreferrer noopener">10.1016/s0166-3542(99)00042-x</a>
An N-terminal arginine-rich cluster and a proline-alanine-threonine repeat region determine the cellular localization of the herpes simplex virus type 1 ICP34.5 protein and its ligand, protein phosphatase 1.
*Repetitive Sequences; Alanine/metabolism; Amino Acid; Amino Acid Sequence; Animals; Arginine/metabolism; Base Sequence; Cell Compartmentation; Cercopithecus aethiops; DNA Primers; Fluorescent Antibody Technique; Indirect; Ligands; Molecular Sequence Data; Phosphoprotein Phosphatases/*metabolism; Proline/metabolism; Protein Phosphatase 1; Recombinant Proteins/chemistry/metabolism; Subcellular Fractions/metabolism; Threonine/metabolism; Vero Cells; Viral Proteins/chemistry/*metabolism
The ICP34.5 protein facilitates herpes simplex virus replication by binding and activating protein phosphatase 1 (PP1) by means of a very conserved C-terminal
Mao Hanwen; Rosenthal Kenneth S
The Journal of biological chemistry
2002
2002-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1074/jbc.M111553200" target="_blank" rel="noreferrer noopener">10.1074/jbc.M111553200</a>
Mild acidic pH inhibition of the major pathway of herpes simplex virus entry into HEp-2 cells.
Animals; Cell Line; Electron; Electrophoresis; Endocytosis; Glycoproteins/analysis; Glycosylation; Humans; Hydrogen-Ion Concentration; Microscopy; Polyacrylamide Gel; Simplexvirus/*physiology/ultrastructure; Temperature; Time Factors; Vero Cells; Viral Proteins/analysis/metabolism
Penetration of the KOS strain of herpes simplex virus type 1 (HSV-1) and the MS and 333 strains of herpes simplex virus type 2 (HSV-2) into HEp-2 cells at pH 6.3 was at least 100-fold less efficient than at pH 7.4. Penetration of two low passage clinical isolates was completely blocked at pH 6.3. The syncytium-forming
Rosenthal K S; Killius J; Hodnichak C M; Venetta T M; Gyurgyik L; Janiga K
The Journal of general virology
1989
1989-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1099/0022-1317-70-4-857" target="_blank" rel="noreferrer noopener">10.1099/0022-1317-70-4-857</a>
Strain-dependent structural variants of herpes simplex virus type 1 ICP34.5 determine viral plaque size, efficiency of glycoprotein processing, and viral release and neuroinvasive disease potential.
*Genetic Variation; Animals; Cercopithecus aethiops; Cultured; Encephalitis; Genetic; Herpes Simplex/*physiopathology/virology; Herpesvirus 1; Human/*classification/genetics/*pathogenicity; Inbred BALB C; Male; Mice; Recombination; Transfection; Tumor Cells; Vero Cells; Viral Envelope Proteins/metabolism; Viral Plaque Assay; Viral Proteins/*chemistry/*genetics/metabolism
The ability of certain strains of herpes simplex virus type 1 (HSV-1) to cause encephalitis or neuroinvasive disease in the mouse upon peripheral infection is dependent on a combination of activities of specific forms of viral proteins. The importance of specific variants of ICP34.5 to neuroinvasive disease potential and its correlation with small-plaque production, inefficient glycoprotein processing, and virus release were suggested by comparison of ICP34.5 from the SP7 virus, originally obtained from the brain of a neonate with disseminated disease, and the tissue culture-passaged progeny of SP7 (SLP5 and SLP10) and the KOS321 virus. SLP5, SLP10, and KOS321 are attenuated and exhibit a large-plaque phenotype, including efficient glycoprotein processing and viral release. We show that expression of the KOS321 ICP34.5 protein in cells infected with SP7 or ICP34.5 deletion mutants promotes large plaque formation and efficient viral glycoprotein processing, while expression of the SP7 ICP34.5 protein decreases efficiency of viral glycoprotein processing. In addition, a recombinant virus, 4hS1, with the SP7 ICP34.5 gene replacing the KOS321-like ICP34.5 gene in the SLP10a background, rescues the small-plaque phenotype and neuroinvasive disease. The major difference in the ICP34.5 gene product is the number of Pro-Ala-Thr repeats in the middle region of the protein, with 18 for SP7 and 3 for KOS321. Strain-dependent differences in the ICP34.5 protein can therefore alter the tissue culture behavior and the virulence of HSV-1.
Mao Hanwen; Rosenthal Ken S
Journal of virology
2003
2003-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1128/jvi.77.6.3409-3417.2003" target="_blank" rel="noreferrer noopener">10.1128/jvi.77.6.3409-3417.2003</a>
Synthesis of radioactive single-stranded DNA probes using asymmetrical PCR and oligonucleotide random priming.
Animals; Isotope Labeling; Cercopithecus aethiops; Vero Cells; DNA; Deoxyribonuclease EcoRI; *Oligonucleotides; *Phosphorus Radioisotopes; DNA Probes/*chemical synthesis; Nucleic Acid Denaturation; Polymerase Chain Reaction/*methods; Northern; Blotting; RNA; Messenger/analysis; Genetic; Herpesvirus 1; *DNA; Viral/analysis; Human/genetics; Single-Stranded; Templates
Mao H; Rosenthal K S
BioTechniques
1999
1999-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.2144/99274bm08" target="_blank" rel="noreferrer noopener">10.2144/99274bm08</a>
Tromantadine inhibits a late step in herpes simplex virus type 1 replication and syncytium formation.
Time Factors; Animals; Rabbits; Fluorescent Antibody Technique; Viral Envelope Proteins/biosynthesis; Virus Replication/*drug effects; Vero Cells; Antibodies; Amantadine/*analogs & derivatives/pharmacology; Cycloheximide/pharmacology; Giant Cells/*drug effects; Glycoproteins/biosynthesis; Membrane Proteins/biosynthesis; Neutralization Tests; Simplexvirus/*drug effects/physiology; Viral/immunology
Addition of tromantadine after virus penetration inhibited HSV-1 induced syncytium formation and virus production in HEp-2 and VERO cells and acted additively with neutralizing antibody in blocking virus spread and cytopathology. Inhibition of syncytium formation in VERO cells infected with 0.01 pfu/cell of
Ickes D E; Venetta T M; Phonphok Y; Rosenthal K S
Antiviral research
1990
1990-08
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/0166-3542(90)90045-9" target="_blank" rel="noreferrer noopener">10.1016/0166-3542(90)90045-9</a>
Herpes simplex virus type 1 penetration initiates mobilization of cell surface proteins.
Animals; Hydrogen-Ion Concentration; In Vitro Techniques; Actin Cytoskeleton/physiology; Vero Cells; Microscopy; Endocytosis; Amantadine/analogs & derivatives/pharmacology; Colchicine/pharmacology; Cytochalasin B/pharmacology; Herpes Simplex/*physiopathology; Membrane Fluidity; Membrane Proteins/*physiology; Microtubules/physiology; Simplexvirus/*physiology; Fluorescence
Changes in membrane structure resulting from herpes simplex virus 1 (HSV-1) penetration were detected using fluorescence photobleaching recovery methods. The effect could be blocked by inhibitors of viral and cellular processes involved in virus penetration. A rapid mode of HSV-1 strain KOS penetration into VERO cells at 37 degrees C normally occurs after a 5 min lag period and is 90-95% complete within 20-30 min. Rates of cell surface protein diffusion increase 2-3-fold after 5 min and return to normal after 25-30 min, this return correlating temporally with the penetration of the virus. At pH 6.3 the lag period preceeding penetration of HSV is increased to 20 min and penetration proceeds much more slowly than at pH 7.4. Inhibition of virus penetration with cytochalasin B or with the antiherpes drug tromantadine also prevents the HSV-1-induced increase in cell surface protein mobility. Colchicine, which does not block HSV-1 penetration, prevents the recovery of the membrane following virus penetration. Therefore, the changes in membrane structure characterized by increased cell surface protein mobility seem to be caused by virus penetration. Cytoskeletal function and integrity are required for the initiation of, and cell recovery from, virus penetration. A pH-sensitive activity, likely to be a virion fusion glycoprotein, is also required.
Rosenthal K S; Roess D; Barisas B G
Biochimica et biophysica acta
1988
1988-07
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/0005-2736(88)90272-6" target="_blank" rel="noreferrer noopener">10.1016/0005-2736(88)90272-6</a>