Chromosomal and Autoradiographic Studies of Cells Infected with Herpes Simplex Virus

R, Waubke; Hausen, Harald zur; Henle, Werner

doi:10.1128/jvi.2.10.1047-1054.1968

Cited by 41 publications

(3 citation statements)

References 13 publications

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“…The described association with condensed chromatin or chromosomes were chance observations, because the parental DNA was localized just as frequently in the nucleoplasm without any particular association with condensed chromatin material. These results confirm published observations by light microscopy in which it was found that most of the viral DNA remains unassociated with chromosomes and is not detectable at sites of chromosomal lesions (12).…”

Section: Discussionsupporting

confidence: 92%

Early Events in Herpes Simplex Virus Infection: a Radioautographic Study

Hummeler

Tomassini

Zając

1969

J Virol

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The early events in herpes simplex virus infection were studied by means of radioautography. The virus was rapidly taken up by the host cells and uncoated. Viral deoxyribonucleic acid (DNA) reached the nuclear sites of replication in 15 to 30 min after infection. The viral DNA occasionally associated with chromosomes or condensed chromatin but was more frequently found to be randomly distributed. Viral progeny appeared 3 hr after infection. These particles did not show any particular spatial relationship to the parental DNA. The morphological latent period lasted 2.5 hr.Morphological studies on the entry of herpes simplex virus (HSV) into tissue culture cells and subsequent early intracellular events have been described (3-5, 7, 8, 10). During this early phase of the infection process, the virus can be followed until the genome is uncoated. The actual transport of the genome to the nucleus, the site of virus replication, cannot be seen. The present study therefore was undertaken to determine the path of the viral deoxyribonucleic acid (DNA) from cell entry to appearance of viral progeny in the nucleus by means of radioautography. This permitted the determination of the length of time required for the viral DNA to reach the nucleus, the intranuclear sites of replication, and spatial relationships of viral progeny to the parental DNA. MATERIALS AND METHODSTissue culture. A BHK-21 line of Syrian hamster kidney cells was grown as monolayers in Blake bottles in a previously described medium (9), which consisted of a mixture of basal medium Eagle (2X) in Hanks balanced salt solution, 10% fetal calf serum, and 10% Tryptose phosphate broth.Virus. The H strain of HSV was received through the courtesy of T. Tokumaru. The virus had been propagated in primary rabbit kidney cells. Before use in these experiments, it was passed five times in BHK-21 cells.Infectivity assay. Plastic petri dishes (60 mm) were seeded with BHK-21 cells in the above medium. They were incubated at 37 C in a 5% CO2 atmosphere. Confluent monolayers were apparent after 24 hr. At this time, the medium was drained and the cells were inoculated with 0.2 ml of the various dilutions of virus.After an adsorption period of 1 hr at 37 C, with occasional agitation, the infected monolayers of cells were overlaid with 5 ml of medium containing 0.5% Difco agar. Cultures were incubated for 72 hr in CO2 atmosphere. After incubation, the overlay medium was removed, and the cells were fixed in 10% neutral Formalin and stained with Leishman's stain (9). The apparent plaques were enumerated and titers were expressed as number of plaque forming units (PFU) per ml.Preparation of labeled virus. Monolayer cultures of 108 BHK-21 cells were infected with HSV at an input multiplicity of 10. At the time of infection, 25 uc/ml of 3H-thymidine per ml (Schwarz BioResearch Inc.), with a specific activity of 6.0 c/mmole, was added to the virus-containing medium. The cultures were incubated at 37 C for 18 to 20 hr when cytopathic effect was evident. The medium was discarded and the ce...

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Section: Discussionsupporting

confidence: 92%

Early Events in Herpes Simplex Virus Infection: a Radioautographic Study

Hummeler

Tomassini

Zając

1969

J Virol

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“…In vitro, a variety of herpes simplex virus (HSV)-induced effects on the host cell genome have been described. HSV has been shown to induce chromosomal aberrations (8,22,62) and host cell DNA repair (43). HSV induces point mutations within the host cell genome, with an efficiency similar to that of chemical carcinogens (49,50,59).…”

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confidence: 99%

A subset of herpes simplex virus replication genes induces DNA amplification within the host cell genome

Heilbronn

Hausen

1989

J Virol

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Herpes simplex virus (HSV) induces DNA amplification of target genes within the host cell chromosome. To characterize the HSV genes that mediate the amplification effect, combinations of cloned DNA fragments covering the entire HSV genome were transiently transfected into simian virus 40 (SV40)-transformed hamster cells. This led to amplification of the integrated SV40 DNA sequences to a degree comparable to that observed after transfection of intact virion DNA. Transfection of combinations of subclones and of human cytomegalovirus immediate-early promoter-driven expression constructs for individual open reading frames led to the identification of six HSV genes which together were necessary and sufficient for the induction of DNA amplification: UL30 (DNA polymerase), UL29 (major DNA-binding protein), UL5, UL8, UL42, and UL52. All of these genes encode proteins necessary for HSV DNA replication. However, an additional gene coding for an HSV origin-binding protein (UL9) was required for origin-dependent HSV DNA replication but was dispensible for SV40 DNA amplification. Our results show that a subset of HSV replication genes is sufficient for the induction of DNA amplification. This opens the possibility that HSV expresses functions sufficient for DNA amplification but separate from those responsible for lytic viral growth. HSV infection may thereby induce DNA amplification within the host cell genome without killing the host by lytic viral growth. This may lead to persistence of a cell with a new genetic phenotype, which would have implications for the pathogenicity of the virus in vivo.

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“…Our findings are also in agreement with those of studies in which breakage has been investigated in cells infected in vitro with herpes sitnplex viruses. Increased breakage and other aberrations have been found in Chinese hamster cells (11,12), in monkey kidney cells (13), in human cell lines HeLa and Hep-2 (14), in human embryo kidney cells (15), in human leukocytes (16,17), and in human diploid fibroblasts (18); also, in human embryo lung cells (15,19) and in human embryo kidney cells and leukocytes (15) after infection with herpes simplex virus Type 2 (HSV-2). On the contrary, in a few studies no damage has been found in human embryo lung cells and in phytohaemagglutinin-stimulated peripheral blood leukocytes aftter infection with HSV-1 (15).…”

Section: Discussionmentioning

confidence: 99%

Chromosomal breaks and sister chromatid exchanges (SCE) in patients with herpetic stomatitis

Zervou-Valvi

Bazopoulou-Kyrkanidou

Angelopoulos

1986

J Oral Pathology Medicine

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Zervou-Valvi F, Biizopoulou-Kyrkanidou E, Angelopoulos AP. Chromosomal breaks and sister chtomatid exchanges (SCE) in patients with herpetie stomatitis. .) Oral Pathol 1986: 15: LS 1-154.Chromosomal breakage and sister-chroniatid exchanges (SCE) were studied in peripheral lymphocytes of 9 patients with primary herpetie stomatitis (PHS). 12 patients with secondary herpetie stomatitis (SnS) and 12 controls. The incidence of chromosomal breakage was signifieantly higher in PHS patients (mean 23%, P.=().0()()2) and in SHS patients (mean 20.25%, P. = 0.0003) compared to the controls (mean 4.2%). The incidence of SCE per 46 chromosomes was significantly higher in SHS patients (mean 16.564) compared with (P.

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Chromosomal and Autoradiographic Studies of Cells Infected with Herpes Simplex Virus

Abstract: The induction of chromosomal aberrations by herpes simplex virus (HSV) and the interaction between viral deoxyribonucleic acid (DNA) and chromosomes have

Cited by 41 publications

References 13 publications

Early Events in Herpes Simplex Virus Infection: a Radioautographic Study

Early Events in Herpes Simplex Virus Infection: a Radioautographic Study

A subset of herpes simplex virus replication genes induces DNA amplification within the host cell genome

Chromosomal breaks and sister chromatid exchanges (SCE) in patients with herpetic stomatitis

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