RNA Polymerase Activity and Inhibition in Herpesvirus-Infected Cells

Sasaki, Yukiko; Sasaki, Ryuzo; Cohen, Gary H.; Pizer, Lewis I.

doi:10.1159/000149751

Cited by 8 publications

(4 citation statements)

References 20 publications

(26 reference statements)

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“…Cells and virus. The procedures for growth of cultured cells, preparation of virus stocks, and plaque assay of virus have been previously described (57,59). The virus, HSV-1 (KOS), was grown and assayed on a continuous line of rabbit skin cells.…”

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confidence: 99%

ICP4-binding sites in the promoter and coding regions of the herpes simplex virus gD gene contribute to activation of in vitro transcription by ICP4

Tedder

Everett

Wilcox

et al. 1989

J Virol

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The herpes simplex virus immediate-early gene product ICP4 activates the transcription of viral early and late genes. We characterized the DNA sequence elements of the early glycoprotein D (gD) gene that play a role in the response to ICP4 in vitro. Using gel mobility shift assays and DNase I footprinting, we identified three ICP4-binding sites, two 5' to the mRNA start site and a third within the coding region. Site II, which gave a footprint between nucleotides-75 and-111 relative to the RNA start site, was previously identified by Faber and Wilcox and contained the reported consensus ICP4-binding site. Site III, which was located between nucleotides + 122 and +163, was very similar to the site II sequence, including a core consensus binding sequence, TCGTC. The site I sequence (nucleotides-308 to-282), however, did not share significant homology with either site II or site III. In vitro transcription experiments from mutant constructs of the gD promoter indicated that all three ICP4-binding sites contribute to the stimulation of transcription by ICP4. DNase I footprinting of the gD promoter with uninfected nuclear extracts of HeLa cells showed protection of two very G-rich sequences between nucleotides-33 and-75. We propose that optimal transcription of the gD gene depends on the interaction of ICP4 with multiple binding sites across the gene and cellular factors that recognize specific sequence elements in the promoter.

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confidence: 99%

ICP4-binding sites in the promoter and coding regions of the herpes simplex virus gD gene contribute to activation of in vitro transcription by ICP4

Tedder

Everett

Wilcox

et al. 1989

J Virol

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“…42,1982 on October 3, 2020 by guest http://jvi.asm.org/ Downloaded from polymerase or one of its activating factors. The search for new or altered polymerase activities has been unsuccessful (23,26,34,39). However, most of these studies have involved the purification of polymerases under conditions where any HSV-specific regulatory subunits could have become dissociated from the enzyme complex.…”

Section: D)iscussionmentioning

confidence: 99%

Herpes simplex virus-induced changes in cellular and adenovirus RNA metabolism in an adenovirus type 5-transformed human cell line

Stenberg¹,

Pizer²

1982

J Virol

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We used the viral transcripts (designated Ad-RNA) that accumulated in the cytoplasm of adenovirus type 5-transformed human embryonic kidney cells (cell line 293-31) as models for cellular RNAs to examine how herpes simplex virus modifies cellular RNA metabolism. Infection of 293-31 cells with herpes simplex virus type 1 strain 17 led to extensive inhibition of Ad-RNA accumulation by 4 h postinfection. The major part of this inhibition was due to an immediate early or alpha gene function, which reduced the rate of transcription of Ad-RNA within the nuclei of the infected cells. In addition, host polyadenylic acid-containing RNA accumulation and rRNA accumulation were affected, but to a lesser extent and at a slower rate than Ad-RNA accumulation. In conjunction with previous data, our experimental data allowed us to propose a general scheme for how herpes simplex virus type 1 alters the metabolism of cellular RNA, the possible mechanisms for these changes, and how they correlate with the regulation of herpes simplex virus gene expression.

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“…RNA polymerase II showed very low concentration of Mg2+ and was inhibited by more than 10 mM Mg2+, though nucleus RNA polymerase from tobacco leaf, which was not separated into polymerase I and II, required 8 mM for maximum activity.I3) Furthermore, other plant RNA polymerase lIs showed maximum activity at 10", 30 mM Mg2+ and still ramained active at 40 mM. I , [4][5][6][7][8][9] RNA polymerase II transcribed denatured calf thymus DNA better than a native calf thymus DNA as template.…”

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confidence: 99%

“…Basic composition of the medium was the same as Linsmaier and Skoog's.m Three day-old cells transferred into the medium containing 10-0 M 2,4-D were harvested (termed 3D-D6 cells) and stored in the deep freezer as the source of enzymes. RNA polymerase preparation was essentially according to the methods described by Sasaki et al 9 Agr. BioI.…”

mentioning

confidence: 99%