The mammalian upstream element factor recognizes two sites in the adenovirus type 2 IVa2-major late promoter intergenic region and stimulates both promoters

Moncollin, Vincent; Kempf, Ashley; Egly, Jean‐Marc

doi:10.1128/jvi.64.7.3199-3206.1990

Cited by 11 publications

(11 citation statements)

References 39 publications

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“…(Figure 2A, lanes 6 and 12). This result was exactly identical to what was observed with the UEFh (20). Both complexes can be competed out by the MLP-UE wild-type oligonucleotide, but neither by the MLP-UE mutant nor by poly(dIdC) (data not shown).…”

Section: Purification Of a Yeast Homolog Of The Hela Uefsupporting

confidence: 87%

“…The binding of this protein to the MLP-UE produces a 3 to 5-fold stimulation of MLP in a HeLa in vitro transcription system. This factor which has been purified from HeLa cells (5,18,19,20) has also been detected in lymphocytes (our unpublished results). A yeast centromere binding protein was also found to recognize sequences homologous to the MLP-UE (21,22).…”

Section: Introductionmentioning

confidence: 64%

“…Protein were analysed by electrophoresis on 9% SDSpolyacrylamide gels. The UEFh was purified by the DNA affinity column using the same procedure as above but starting with the SPO.35 fraction prepared as described previously (5,20). The purified UEFh fraction (5 jig/ml) contains two polypeptides of 43 and 45 kD (20).…”

Section: Purification Of the Yeast Uefmentioning

confidence: 99%

“…UEFh; see 5,18,19,20). The yeast and human UEF interact with identical DNA sequences In order to precisely delineate the DNA region that interacts with the yeast protein, DNAse I footprint experiment was performed.…”

Section: Purification Of a Yeast Homolog Of The Hela Uefmentioning

confidence: 99%

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A yeast homolog of the human UEF stimulates transcription from the adenovirus 2 major late promoter in yeast and in mammalian cell-free systems

Moncollin¹,

Stalder²,

Verdier³

et al. 1990

Nucl Acids Res

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We report the identification and purification of a yeast factor functionally homologous to the human upstream element factor (UEFh). Although the yeast protein (UEFy) has a higher molecular weight than the HeLa UEF (60 kD versus 45 kD) both have identical DNA-binding properties: the purified UEFy recognizes the Adenovirus 2 (Ad2) major late promoter upstream element (MLP-UE; from nucleotide -49 to -67) as well as the IVa2 upstream element (IVa2-UE; from nucleotide -98 to -122) with a higher affinity for the MLP-UE than for the IVa2-UE. Based on its DNA binding specificity, size and thermostability, the UEFy protein appears also similar or equivalent to the centromere binding protein CP1. In a competition assay with oligonucleotides containing the MLP-UE binding site, a drastic reduction of Ad2 MLP transcription was observed both in a HeLa and in a yeast cell free system, which was restored by addition of either the purified UEFh or UEFy proteins. We conclude that both UEFh and UEFy activate transcription from the Ad2 MLP upon binding to the upstream element, whatever is the in vitro cell-free system (yeast or HeLa). This indicate that some regulatory function represented by the upstream element and its cognate factor, is well conserved between human and yeast.

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Section: Purification Of a Yeast Homolog Of The Hela Uefsupporting

confidence: 87%

Section: Introductionmentioning

confidence: 64%

Section: Purification Of the Yeast Uefmentioning

confidence: 99%

Section: Purification Of a Yeast Homolog Of The Hela Uefmentioning

confidence: 99%

See 2 more Smart Citations

A yeast homolog of the human UEF stimulates transcription from the adenovirus 2 major late promoter in yeast and in mammalian cell-free systems

Moncollin¹,

Stalder²,

Verdier³

et al. 1990

Nucl Acids Res

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“…1). This arrangement of divergent transcription units, in conjunction with results of in vitro transcription and DNase I footprinting assays, led to the proposal that the ML and the IVa 2 promoters share binding sites for USF/MLTF and thus compete for this transcriptional regulator (Adami and Babiss, 1992;Carcamo et al, 1989;Moncollin et al, 1990;Natarajan et al, 1984). In principle, such competition could preclude IVa 2 transcription during the early phase of infection, when ML transcription is stimulated by the 289R E1A protein (Leong et al, 1988;Nevins, 1981).…”

Section: Introductionmentioning

confidence: 99%

Identification of a Cellular Repressor of Transcription of the Adenoviral Late IVa2 Gene That Is Unaltered in Activity in Infected Cells

Lin

Flint

2000

Virology

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The gene encoding the adenovirus type 2 IVa(2) protein, a sequence-specific activator of transcription from the viral major late promoter, is itself transcribed only during the late phase of infection. We previously identified a cellular protein (IVa(2)-RF) that binds specifically to an intragenic sequence of the IVa(2) transcription unit. We now report that precise substitutions within the IVa(2)-RF-binding site that decreased binding affinity increased the efficiency of IVa(2) transcription in in vitro reactions containing IVa(2)-RF. Consistent with the conclusion that this cellular protein represses IVa(2) transcription, mutations that led to more efficient transcription in the presence of IVa(2)-RF were without effect in reactions lacking this cellular protein. No change in the concentration or activity of IVa(2)-RF could be detected in adenovirus-infected cells during the period in which the IVa(2) gene is transcribed. We therefore propose that restriction of IVa(2) transcription to the late phase is the result of titration of this cellular repressor as the number of copies of the IVa(2) promoter increases upon replication of the viral genome.

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Transcription from the adenovirus major late promoter uses redundant activating elements.

Reach

Young

1991

The EMBO Journal

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The adenovirus major late promoter (MLP) has been analyzed by constructing recombinant viral genomes containing mutations in possible promoter elements. Single base pair changes in the TATA box had no effect on viral replication, and MLP expression, as measured by the accumulation of late mRNAs, was at wild type levels. However, a double mutation in the TATA box reduced viral replication and MLP expression, demonstrating that the TATA box is important, although not essential, for maximal activity in virus. Primer extension analysis showed that the mRNAs were initiated at the correct position. A mutation in the CAAT box was viable, and had only minor effects on MLP expression. However, this mutation when coupled to a single mutation in the TATA box, severely reduced viral replication and expression from the MLP. Similarly, a viable mutation in the UPE, shown previously to abolish binding of USF, coupled to a single mutation in the TATA box was lethal. These results suggest that both USF and the CAAT box binding factor CP1 can interact with TFIID to effect activation, and thus that the mechanism of activation is functionally redundant.

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The mammalian upstream element factor recognizes two sites in the adenovirus type 2 IVa2-major late promoter intergenic region and stimulates both promoters

Cited by 11 publications

References 39 publications

A yeast homolog of the human UEF stimulates transcription from the adenovirus 2 major late promoter in yeast and in mammalian cell-free systems

A yeast homolog of the human UEF stimulates transcription from the adenovirus 2 major late promoter in yeast and in mammalian cell-free systems

Identification of a Cellular Repressor of Transcription of the Adenoviral Late IVa2 Gene That Is Unaltered in Activity in Infected Cells

Transcription from the adenovirus major late promoter uses redundant activating elements.

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