scholarly journals A mutant herpesvirus protein leads to a block in nuclear localization of other viral proteins.

1986 ◽  
Vol 6 (7) ◽  
pp. 2371-2381 ◽  
Author(s):  
D M Knipe ◽  
J L Smith
Keyword(s):  
Herpes Simplex ◽  
Nuclear Localization ◽  
Cell Nucleus ◽  
Viral Proteins ◽  
The Other ◽  
Temperature Sensitive ◽  
Nonpermissive Temperature ◽  
Primary Defect ◽  
Infected Cells ◽  
Simplex Virus

The herpes simplex virus mutants KOS1.1 ts756 and HFEM tsLB2 express temperature-sensitive ICP4 proteins that are not localized properly to the cell nucleus at the nonpermissive temperature. In these infected cells at the nonpermissive temperature, nuclear localization of at least two other viral proteins, ICP0 and ICP8, is impaired. Replacement of the mutated sequences in the ICP4 gene of tsLB2 restored proper nuclear localization of all of the proteins. The ICP0 and ICP8 proteins expressed in cells transfected with their individual genes were localized to the cell nucleus. Therefore, in infected cells, the mutant ICP4 gene product appears to be the primary defect which leads to the block in nuclear localization of the other proteins. One viral protein, ICP27, was not inhibited for nuclear localization in these cells. These data indicate that there are at least two pathways for nuclear localization of HSV proteins, one of which is inhibited by the mutant ICP4 protein. The mutant ICP4 protein may define a probe for one of the pathways of nuclear localization of proteins.

A mutant herpesvirus protein leads to a block in nuclear localization of other viral proteins

1986 ◽  
Vol 6 (7) ◽  
pp. 2371-2381
Author(s):  
D M Knipe ◽  
J L Smith
Keyword(s):  
Herpes Simplex ◽  
Nuclear Localization ◽  
Cell Nucleus ◽  
Viral Proteins ◽  
The Other ◽  
Temperature Sensitive ◽  
Nonpermissive Temperature ◽  
Primary Defect ◽  
Infected Cells ◽  
Simplex Virus

The herpes simplex virus mutants KOS1.1 ts756 and HFEM tsLB2 express temperature-sensitive ICP4 proteins that are not localized properly to the cell nucleus at the nonpermissive temperature. In these infected cells at the nonpermissive temperature, nuclear localization of at least two other viral proteins, ICP0 and ICP8, is impaired. Replacement of the mutated sequences in the ICP4 gene of tsLB2 restored proper nuclear localization of all of the proteins. The ICP0 and ICP8 proteins expressed in cells transfected with their individual genes were localized to the cell nucleus. Therefore, in infected cells, the mutant ICP4 gene product appears to be the primary defect which leads to the block in nuclear localization of the other proteins. One viral protein, ICP27, was not inhibited for nuclear localization in these cells. These data indicate that there are at least two pathways for nuclear localization of HSV proteins, one of which is inhibited by the mutant ICP4 protein. The mutant ICP4 protein may define a probe for one of the pathways of nuclear localization of proteins.

scholarly journals Effects of Major Capsid Proteins, Capsid Assembly, and DNA Cleavage/Packaging on the pUL17/pUL25 Complex of Herpes Simplex Virus 1

2009 ◽  
Vol 83 (24) ◽  
pp. 12725-12737 ◽  
Author(s):  
Luella Scholtes ◽  
Joel D. Baines
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Nuclear Localization ◽  
Conformational Changes ◽  
Viral Proteins ◽  
Dna Packaging ◽  
Capsid Proteins ◽  
Herpes Simplex Virus 1 ◽  
Infected Cells ◽  
Simplex Virus

ABSTRACT The UL17 and UL25 proteins (pUL17 and pUL25, respectively) of herpes simplex virus 1 are located at the external surface of capsids and are essential for DNA packaging and DNA retention in the capsid, respectively. The current studies were undertaken to determine whether DNA packaging or capsid assembly affected the pUL17/pUL25 interaction. We found that pUL17 and pUL25 coimmunoprecipitated from cells infected with wild-type virus, whereas the major capsid protein VP5 (encoded by the UL19 gene) did not coimmunoprecipitate with these proteins under stringent conditions. In addition, pUL17 (i) coimmunoprecipitated with pUL25 in the absence of other viral proteins, (ii) coimmunoprecipitated with pUL25 from lysates of infected cells in the presence or absence of VP5, (iii) did not coimmunoprecipitate efficiently with pUL25 in the absence of the triplex protein VP23 (encoded by the UL18 gene), (iv) required pUL25 for proper solubilization and localization within the viral replication compartment, (v) was essential for the sole nuclear localization of pUL25, and (vi) required capsid proteins VP5 and VP23 for nuclear localization and normal levels of immunoreactivity in an indirect immunofluorescence assay. Proper localization of pUL25 in infected cell nuclei required pUL17, pUL32, and the major capsid proteins VP5 and VP23, but not the DNA packaging protein pUL15. The data suggest that VP23 or triplexes augment the pUL17/pUL25 interaction and that VP23 and VP5 induce conformational changes in pUL17 and pUL25, exposing epitopes that are otherwise partially masked in infected cells. These conformational changes can occur in the absence of DNA packaging. The data indicate that the pUL17/pUL25 complex requires multiple viral proteins and functions for proper localization and biochemical behavior in the infected cell.

scholarly journals Analysis of HCF, the Cellular Cofactor of VP16, in Herpes Simplex Virus-Infected Cells

2000 ◽  
Vol 74 (1) ◽  
pp. 99-109 ◽  
Author(s):  
Sylvie LaBoissière ◽  
Peter O'Hare
Keyword(s):  
Protein Synthesis ◽  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Cell Line ◽  
Temperature Sensitive ◽  
Nonpermissive Temperature ◽  
Wild Type ◽  
Infected Cells ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT Herpes simplex virus (HSV) immediate-early (IE) gene expression is initiated via the recruitment of the structural protein VP16 onto specific sites upstream of each IE gene promoter in a multicomponent complex (TRF.C) that also includes the cellular proteins Oct-1 and HCF. In vitro results have shown that HCF binds directly to VP16 and stabilizes TRF.C. Results from transfection assays have also indicated that HCF is involved in the nuclear import of VP16. However, there have been no reports on the role or the fate of HCF during HSV type 1 (HSV-1) infection. Here we show that the intracellular distribution of HCF is dramatically altered during HSV-1 infection and that the protein interacts with and colocalizes with VP16. Moreover, viral protein synthesis and replication were significantly reduced after infection of a BHK-21-derived temperature-sensitive cell line (tsBN67) which contains a mutant HCF unable to associate with VP16 at the nonpermissive temperature. Intracellular distribution of HCF and of newly synthesized VP16 in tsBN67-infected cells was similar to that observed in Vero cells, suggesting that late in infection the trafficking of both proteins was not dependent on their association. We constructed a stable cell line (tsBN67r) in which the temperature-sensitive phenotype was rescued by using an epitope-tagged wild-type HCF. In HSV-1-infected tsBN67r cells at the nonpermissive temperature, direct binding of HCF to VP16 was observed, but virus protein synthesis and replication were not restored to levels observed at the permissive temperature or in wild-type BHK cells. Together these results indicate that the factors involved in compartmentalization of VP16 alter during infection and that late in infection, VP16 and HCF may have additional roles reflected in their colocalization in replication compartments.

scholarly journals The UL25 Gene Product of Herpes Simplex Virus Type 1 Is Involved in Uncoating of the Viral Genome

2008 ◽  
Vol 82 (13) ◽  
pp. 6654-6666 ◽  
Author(s):  
Valerie G. Preston ◽  
Jill Murray ◽  
Christopher M. Preston ◽  
Iris M. McDougall ◽  
Nigel D. Stow
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Herpes Simplex Virus Type ◽  
Viral Genome ◽  
Dna Packaging ◽  
Nonpermissive Temperature ◽  
Viral Dna ◽  
Infected Cells ◽  
Simplex Virus

ABSTRACT Studies on the herpes simplex virus type 1 UL25-null mutant KUL25NS have shown that the capsid-associated UL25 protein is required at a late stage in the encapsidation of viral DNA. Our previous work on UL25 with the UL25 temperature-sensitive (ts) mutant ts1204 also implicated UL25 in a role at very early times in the virus growth cycle, possibly at the stage of penetration of the host cell. We have reexamined this mutant and discovered that it had an additional ts mutation elsewhere in the genome. The ts1204 UL25 mutation was transferred into wild-type (wt) virus DNA, and the UL25 mutant ts1249 was isolated and characterized to clarify the function of UL25 at the initial stages of virus infection. Indirect immunofluorescence assays and in situ hybridization analysis of virus-infected cells revealed that the mutant ts1249 was not impaired in penetration of the host cell but had an uncoating defect at the nonpermissive temperature. When ts1249-infected cells were incubated initially at the permissive temperature to allow uncoating of the viral genome and subsequently transferred to the restrictive temperature, a DNA-packaging defect was evident. The results suggested that ts1249, like KUL25NS, had a block at a late stage of DNA packaging and that the packaged genome was shorter than the full-length genome. Examination of ts1249 capsids produced at the nonpermissive temperature revealed that, in comparison with wt capsids, they contained reduced amounts of UL25 protein, thereby providing a possible explanation for the failure of ts1249 to package full-length viral DNA.

scholarly journals Herpes Simplex Virus Entry Mediator Associates in Infected Cells in a Complex with Viral Proteins gD and at Least gH

2005 ◽  
Vol 79 (7) ◽  
pp. 4540-4544 ◽  
Author(s):  
Pilar Perez-Romero ◽  
Aleida Perez ◽  
Althea Capul ◽  
Rebecca Montgomery ◽  
A. Oveta Fuller
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Protein Interactions ◽  
Virus Entry ◽  
Viral Proteins ◽  
Receptor Protein ◽  
Infected Cells ◽  
Complex Forms ◽  
Simplex Virus ◽  
In Cells

ABSTRACT We examined herpes simplex virus (HSV)-infected human HEp-2 cells or porcine cells that express herpes virus entry mediator (HVEM) for virus and receptor protein interactions. Antibody to HVEM, or its viral ligand gD, coimmunoprecipitated several similar proteins. A prominent 110-kDa protein that coprecipitated was identified as gH. The HVEM/gD/gH complex was detected with mild or stringent cell lysis conditions. It did not form in cells infected with HSV-1(KOS)Rid1 virus or with null virus lacking gD, gH, or gL. Thus, in cells a complex forms through physical associations of HVEM, gD, and at least gH.

Cell-mediated immunity to herpes simplex virus: specificity of cytotoxic T cells

1980 ◽  
Vol 30 (2) ◽  
pp. 451-461
Author(s):  
M J Lawman ◽  
R J Courtney ◽  
R Eberle ◽  
P A Schaffer ◽  
M K O'Hara ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Target Cells ◽  
Temperature Sensitive ◽  
Nonpermissive Temperature ◽  
Binding Studies ◽  
Glycoprotein Synthesis ◽  
Viral Antigens ◽  
Simplex Virus ◽  
Hsv 1

This communication deals with the question of which of the viral antigens constitutes the targets for cytotoxic T lymphocytes (CTL) generated against herpes simplex virus type 1 (HSV-1). The approach used was, first, to compare cytotoxicity of CTL against target cells infected with virus in the presence of tunicamycin and 2-deoxy-D-glucose, which are known to inhibit glycoprotein synthesis, and second, to compare cytotoxicity of CTL against target cells infected with wild-type HSV-1 with that against target cells infected with a temperature-sensitive mutant of HSV-1 which, at the nonpermissive temperature, exhibits diminished glycoprotein synthesis. The results show that glycoprotein expression is required for the demonstration of cytotoxic activity of CTL. The level of cytotoxicity against the temperature-sensitive HSV-1 target at the nonpermissive temperature was reduced and correlated with the level of expression of the major envelope glycoprotein region (VP123; molecular weight = 123,000) at the target cell surface as measured serologically by antibody binding studies. The results were interpreted to indicate that HSV-1-induced glycoproteins are the target antigens for anti-HSV CTL and that the principal viral antigens recognized by the CTL may be glycoproteins of the VP123 region.

scholarly journals Temperature-Sensitive Mutations in the Putative Herpes Simplex Virus Type 1 Terminase Subunits pUL15 and pUL33 Preclude Viral DNA Cleavage/Packaging and Interaction with pUL28 at the Nonpermissive Temperature

2007 ◽  
Vol 82 (1) ◽  
pp. 487-494 ◽  
Author(s):  
Kui Yang ◽  
Alice P. W. Poon ◽  
Bernard Roizman ◽  
Joel D. Baines
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Virus Type ◽  
Herpes Simplex Virus Type ◽  
Dna Cleavage ◽  
Temperature Sensitive ◽  
Nonpermissive Temperature ◽  
Viral Dna ◽  
Simplex Virus

ABSTRACT Terminases comprise essential components of molecular motors required to package viral DNA into capsids in a variety of DNA virus systems. Previous studies indicated that the herpes simplex virus type 1 UL15 protein (pUL15) interacts with the pUL28 moiety of a pUL28-pUL33 complex to form the likely viral terminase. In the current study, a novel temperature-sensitive mutant virus was shown to contain a mutation in UL33 codon 61 predicted to change threonine to proline. At the nonpermissive temperature, this virus, designated ts8-22, replicated viral DNA and produced capsids that became enveloped at the inner nuclear membrane but failed to form plaques or to cleave or package viral DNA. Incubation at the nonpermissive temperature also precluded coimmunoprecipitation of UL33 protein with its normal interaction partners encoded by UL28 and UL15 in ts8-22-infected cells and with pUL28 in transient-expression assays. Moreover, a temperature-sensitive mutation in UL15 precluded coimmunoprecipitation of pUL15 with the UL28 and UL33 proteins at the nonpermissive temperature. We conclude that interactions between putative terminase components are tightly linked to successful viral DNA cleavage and packaging.

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