A Filamentous Distribution for the Herpes Simplex Virus Type 2-encoded Major DNA-binding Protein

Rose, Dietlind; Shriver, K; Latchman, David S.; LaThangue, N. B.

doi:10.1099/0022-1317-67-7-1315

Cited by 11 publications

(3 citation statements)

References 37 publications

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“…The ICP8 MAB, 39S (Showalter et al, 1981), was prepared from ascites fluid of animals inoculated with hybridoma cells derived from cultures originally obtained from ATCC and was used at a 1:25 dilution. The ICP8 MAB, 10E3 (Rose et al, 1986), obtained from Kathleen Shriver was used at a 1:40 dilution. The ICP4 MAB, 58S, (Showalter et al, 1981) was a gift from Neal DeLuca, Univ.…”

Section: Antibodies and Reagentsmentioning

confidence: 99%

Comparison of the Intranuclear Distributions of Herpes Simplex Virus Proteins Involved in Various Viral Functions

et al. 1998

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Herpesviral transcription, DNA synthesis, and capsid assembly occur within the infected cell nucleus. To further define the spatial relationship among these processes, we have examined the intranuclear distributions of viral DNA replication, gene regulatory, and capsid proteins using dual label immunofluorescence and confocal microscopy. We observed that several of the viral DNA replication proteins localize preferentially to punctate structures within replication compartments while the major transcriptional activator, ICP4, and the ICP27 regulatory protein show a more diffuse distribution within replication compartments. The viral proteins that show a punctate distribution in replication compartments redistribute from these compartments to prereplicative sites when viral DNA replication is inhibited, whereas viral proteins that show a diffuse distribution remain within replication compartments when viral DNA replication is inhibited. Thus the sites of viral DNA replication and late transcription appear to be distinct but codistribute within the boundaries of replication compartments. The major capsid protein, ICP5, also localizes initially to a diffuse distribution within replication compartments, but during the time of maximal progeny virus assembly, ICP5 becomes localized to punctate structures within replication compartments that are often near the punctate structures occupied by viral DNA replication proteins. Hence the processes of viral DNA replication, late transcription, and capsid assembly show a general overlapping distribution within replication compartments but appear to be located at distinct sites within these regions of the infected cell nucleus.

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Section: Antibodies and Reagentsmentioning

confidence: 99%

Comparison of the Intranuclear Distributions of Herpes Simplex Virus Proteins Involved in Various Viral Functions

et al. 1998

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“…5, lane 5), a compound that preferentially inhibits the herpes simplex virus type 1 DNA polymerase. Incorporation of [3H]thymidine into wild-type viral DNA was detected when 1/10 the normal amount of total b The antibody used for the Western blots to visualize ICP8 was rabbit polyclonal 3-83 (24) or mouse monoclonal 1OE-3 (41). The negatives of the color reactions of Westem blots were scanned by densitometer.…”

Section: -mentioning

confidence: 99%

Genetic evidence for multiple nuclear functions of the herpes simplex virus ICP8 DNA-binding protein

Gao

Knipe

1989

J Virol

110

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We have isolated several mutant herpes simplex viruses, specifically mutated in the infected cell protein 8 (ICP8) gene, to define the functional domains of ICP8, the major viral DNA-binding protein. To facilitate the isolation of these mutants, we first isolated a mutant virus, HD-2, with the lacZ gene fused to the ICP8 gene so that an ICP8-40-galactosidase fusion protein was expressed. This virus formed blue plaques on ICP8-expressing cell lines in the presence of 5-bromo-4-chloro-3-indolyl-,1-D-galactopyranoside. Mutated ICP8 gene plasmids contransfected with HD-2 DNA yielded recombinant viruses with the mutant ICP8 gene incorporated into the viral genome. These recombinants were identified by formation of white plaques. Four classes of mutants were defined: (i) some expressed ICP8 that could bind to DNA but could not localize to the cell nucleus; (ii) some expressed ICP8 that did not bind to DNA but localized to the nucleus; (iii) some expressed ICP8 that neither bound to DNA nor localized to the nucleus; and (iv) one expressed ICP8 that localized to the cell nucleus and bound to DNA in vitro, but the mutant virus did not replicate its DNA. These classes of mutants provide genetic evidence that DNA binding and nuclear localization are distinct functions of ICP8 and that ICP8 has nuclear functions other than binding to DNA. Furthermore, the portion of ICP8 needed for a nuclear function(s) distinct from DNA binding is the part of ICP8 showing sequence similarity to that of the cellular protein cyclin or proliferating cell nuclear antigen. MATERIALS AND METHODS Cells and viruses. Vero cells were grown and maintained as described previously (23). The growth medium for the Neor cell lines S2 and B10 (see below) included 200 ,ug of the antibiotic G418 per ml during the first passage of cells after thawing or 500 pg of G418 per ml every five passages. The herpes simplex virus type 1 wild-type strain KOS1.1 was propagated and assayed as described previously (23, 25). Mutant viruses were grown in ICP8-expressing S2 and 5258

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“…1) (42) as described previously (6,10). After growth in medium containing the antibiotic G418 (a neomycin analog), drugresistant colonies were isolated, grown into cultures, and screened by indirect immunofluorescence (5) using the lOE-3 anti-ICP8 monoclonal antibody (38) for the ability to express the mutant form of ICP8 upon nlO infection. The lOE-3 antibody recognizes diO5 ICP8 but not nlO ICP8 (9b).…”

Section: Methodsmentioning

confidence: 99%

Potential role for herpes simplex virus ICP8 DNA replication protein in stimulation of late gene expression

Gao

Knipe

1991

J Virol

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We have identified a trans-dominant mutant form of the herpes simplex virus (HSV) DNA-binding protein ICP8 which inhibits viral replication. When expressed by the V2.6 cell line, the mutant gene product inhibited wild-type HSV production by 50to 150-fold when the multiplicity of infection was less than 5. Production of HSV types 1 and 2 but not production of pseudorabies virus was inhibited in V2.6 cells. The inhibitory effect was not due solely to the high levels of expression, because the levels of expression were comparable to those in the permissive wild-type ICP8-expressing S-2 cell line. Experiments designed to define the block in viral production in V2.6 cells demonstrated (i) that viral a and I8 gene expression was comparable in the different cell lines, (ii) that viral DNA replication proceeded but was reduced to approximately 20% of the control cell level, and (iii) that late gene expression was similar to that in cells in which viral DNA replication was completely blocked. Genetic experiments indicated that the mutant gene product inhibits normal functions of ICP8. Thus, ICP8 may play distinct roles in replication of viral DNA and in stimulation of late gene expression. The dual roles of ICP8 in these two processes could provide a mechanism for controlling the transition from viral DNA synthesis to late gene expression during the viral growth cycle.

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A Filamentous Distribution for the Herpes Simplex Virus Type 2-encoded Major DNA-binding Protein

Cited by 11 publications

References 37 publications

Comparison of the Intranuclear Distributions of Herpes Simplex Virus Proteins Involved in Various Viral Functions

Comparison of the Intranuclear Distributions of Herpes Simplex Virus Proteins Involved in Various Viral Functions

Genetic evidence for multiple nuclear functions of the herpes simplex virus ICP8 DNA-binding protein

Potential role for herpes simplex virus ICP8 DNA replication protein in stimulation of late gene expression

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