Adenovirus 2 Peptide IX Gene Is Expressed Only on Replicated DNA Molecules

Mizutani, Takashi; Murayama, Minoru; Morita, Takashi

doi:10.1128/mcb.6.12.4149-4154.1986

Cited by 3 publications

(1 citation statement)

References 34 publications

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“…By using the same assay, i.e., transient transfection of HeLa cells, similar results to those presented here for the Xenopus (-globin gene have been obtained for the adenovirus pIX gene (33,47). This gene is expressed in vivo after the onset of viral DNA replication, and superinfection experiments had previously demonstrated that the delayed expression of this gene could not be overcome simply by supplying early gene products in trans (8,43).…”

Section: Discussionsupporting

confidence: 76%

Role for DNA Replication in β-Globin Gene Activation

Enver

Brewer

Patient

1988

Molecular and Cellular Biology

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Transcriptional activation of the Xenopus laevis beta-globin gene requires the synergistic action of the simian virus 40 enhancer and DNA replication in DEAE-dextran-mediated HeLa cell transfections. Replication does not act through covalent modification of the template, since its requirement was not obviated by the prior replication of the transfected DNA in eucaryotic cells. Transfection of DNA over a 100-fold range demonstrates that replication does not contribute to gene activation simply increasing template copy number. Furthermore, in cotransfections of replicating and nonreplicating constructs, only replicating templates were transcribed. Replication is not simply a requirement of chromatin assembly, since even unreplicated templates generated nucleosomal ladders. Stimulation of beta-globin transcription by DNA replication, though less marked, was also observed in calcium phosphate transfections. We interpret these results as revealing a dynamic role for replication in gene activation.

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Section: Discussionsupporting

confidence: 76%

Role for DNA Replication in β-Globin Gene Activation

Enver

Brewer

Patient

1988

Molecular and Cellular Biology

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Identification of a Previously Unrecognized Promoter That Drives Expression of the UXP Transcription Unit in the Human Adenovirus Type 5 Genome

Bai

Tollefson

Wold

2010

J Virol

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We previously identified an adenovirus (Ad) protein named U exon protein (UXP) encoded by a leftward-strand (l-strand) transcription unit. Here we identify and characterize the UXP promoter. Primer extension and RNase protection assays mapped the transcription initiation site at 32 nucleotides upstream of the UXP gene initiation codon. A series of viral mutants with mutations at two putative inverted CCAAT (I-CCAAT) boxes and two E2F sites were generated. With mutants lacking the proximal I-CCAAT box, the UXP mRNA level decreased significantly to 30% of the Ad type 5 (Ad5) mRNA level as measured by quantitative reverse transcription-PCR. Decreased UXP was also observed by immunoblotting and immunofluorescence. UXP mRNA and protein levels were similar to those of Ad5 for mutants lacking the distal I-CCAAT box or both putative E2F sites. Ad DNA levels were similar in mutant-and wild-type Ad5-infected cells during the late stage of infection, strongly suggesting that the decreased UXP mRNA and protein from mutants lacking the proximal I-CCAAT box was due to decreased promoter activity. Electrophoretic mobility shift assays (EMSA) indicated that a cellular factor binds specifically to the proximal I-CCAAT box of the UXP promoter. An in vitro luciferase reporter assay demonstrated that basal promoter activity lies between bp ؊158 and ؉30 of the transcription initiation site. No E1A-mediated promoter transactivation was observed in 293 cells compared with A549 cells. Thus, we propose that there is a previously unidentified Ad5 promoter that drives expression of the UXP transcription unit. This promoter is embedded within the gene for fiber, and it contains a proximal I-CCAAT box critical for UXP mRNA transcription.

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Transcription Factor Access Is Mediated by Accurately Positioned Nucleosomes on the Mouse Mammary Tumor Virus Promoter

Archer

Cordingley

Wolford

et al. 1991

Molecular and Cellular Biology

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A fragment of the mouse mammary tumor virus (MMTV) promoter was reconstituted from pure histones into a dinucleosome with uniquely positioned octamer cores. Core boundaries for the in vitro-assembled dinucleosome corresponded to the observed in vivo phasing pattern for long terminal repeat nucleosomes A and B. Nuclear factor 1 (NF1), a constituent of the MMTV transcription initiation complex, was excluded from the assembled dinucleosome, whereas the glucocorticoid receptor was able to bind. During transcription of MMTV in vivo, displacement of nucleosome B was necessary to permit assembly of the initiation complex. These results indicate that the nucleoprotein structure of the promoter can provide differential access to sequence-specific DNA-binding proteins and that active chromatin remodeling can occur during transcription activation.

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Adenovirus 2 Peptide IX Gene Is Expressed Only on Replicated DNA Molecules

Cited by 3 publications

References 34 publications

Role for DNA Replication in β-Globin Gene Activation

Role for DNA Replication in β-Globin Gene Activation

Identification of a Previously Unrecognized Promoter That Drives Expression of the UXP Transcription Unit in the Human Adenovirus Type 5 Genome

Transcription Factor Access Is Mediated by Accurately Positioned Nucleosomes on the Mouse Mammary Tumor Virus Promoter

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