Analysis of promoters of adenovirus type 12 E1A gene in a cell-free transcription system

Nakanishi, Yoshinobu; Shibata, Hitomi; Hase, Tetsu; Masamune, Yukito

doi:10.1016/0006-291x(87)90598-5

Cited by 9 publications

(4 citation statements)

References 24 publications

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“…Nuclear extracts were prepared from Ehrlich ascites tumor cells as described (33,34). Cell-free transcription was conducted in a 25-,ul reaction mixture containing 12 mM Hepes (pH 7.9), 0.12 mM EDTA, 0.3 mM dithiothreitol, 60 mM KCI, 5 mM MgCl2, 0.3 mM phenylmethylsulfonyl fluoride, 12% (vol/vol) glycerol, 0.5 mM each NTP, 5 mM creatine phosphate, -0.2 mg of extract protein, and the following template DNAs: covalently closed PlHCAT, PlBCAT, and P2EBg at 8 ,g/ ml; P2ECAT, P2PCAT, and pElACAT at 20 ,g/ml; and pHIVCAT at 80 ,g/ml, except that pSmaF DNA was used at 20 ,ug/ml after digestion with Sma I.…”

Section: Methodsmentioning

confidence: 99%

Direct effect of basic fibroblast growth factor on gene transcription in a cell-free system.

Nakanishi

Kihara

Mizuno

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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Recent findings on the translocation of intact fibroblast growth factor (FGF) into the cell nucleus suggest that it functions directly in nuclear events. We examined the effect of human basic FGF (bFGF) on gene transcription in a cell-free system. When mouse genes encoding phosphoglycerate kinases 1 and 2 (Pgk-1 and Pgk-2) were transcribed by using nuclear extracts of Ehrlich ascites tumor cells, FGF affected transcription in different ways: in the presence of bFGF, transcription of the Pgk-1 gene was inhibited, whereas that of the Pgk-2 gene was enhanced. When viral genes were tested, transcription of the adenovirus major late DNA was slightly stimulated but that of the adenovirus early EIA DNA or the human immunodeficiency virus DNA was not changed by the addition of bFGF. Moreover, the presence of a distinct 5' upstream region of the Pgk-2 gene, which includes a negative cis-acting element, was required for transcription stimulation by bFGF. These results suggest that bFGF can regulate transcription directly in the nucleus in a gene-specific manner.

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Section: Methodsmentioning

confidence: 99%

Direct effect of basic fibroblast growth factor on gene transcription in a cell-free system.

Nakanishi

Kihara

Mizuno

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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“…Nuclear extracts were prepared from HeLa and MOLT-4 no. 8 (28) cells as previously described (8,41). The transcription reaction was conducted at 30°C for 1 h in a 25-pI mixture (41) including 0.2 mg of the extracts protein and 2 ,ug of covalently closed DNA as template, a DNA amount which gave maximal transcription (data not shown).…”

mentioning

confidence: 99%

A novel cis-acting element that controls transcription of human immunodeficiency virus type 1 DNA, depending on cell type

Nakanishi

Masamune

Kimura

1991

J Virol

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cis-acting elements for the transcription of human immunodeficiency virus type 1 DNA were analyzed in cell-free transcription and DNA transfection assays. Besides previously identified cis elements, a region adjacent to the enhancer element was found to regulate transcription in both assays. Loss of this region caused 4.3and 1.6-fold transcription inhibition in a transfection assay with a T-cell line, MOLT-4, and a monocyte line, U937, respectively, whereas the same region appeared to function negatively with other T-cell lines, MT-4 and Jurkat. These results suggest that this novel cis element regulates the transcription of proviral DNA in a cell-type-specific manner.

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“…Thus, for Ad12 it was shown by in vitro transcription assays that the promoters of both TS1 and TS2 are individually stimulated by cellular factors Nakanishi et al, 1987). Previous reports have indicated that the binding of nuclear factor-I (NF-I) to a region located near the left end of the Ad12 genome is involved in stimulation of TS1 (Koikeda et al, 1990 ;Shibata et al, 1989) and that the DNA-binding activity of NF-I might be inhibited by the E1A protein (Koikeda et al, 1990 ;Shibata-Sakurai et al, 1991).…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, Ad12 possesses two functional transcriptional start sites located at nt 306 (TS1) and 445 (TS2). Both initiation sites are active at an early stage after infection with virus (Saito et al, 1981 ;Sawada & Fujinaga, 1980), resulting in a long and a short 13S-and 12S-primary transcript (Fujinaga et al, 1984 ;Nakanishi et al, 1987 ;Shibata et al, 1989).…”

Section: Introductionmentioning

confidence: 99%

A cis-acting element 7 bp upstream of the ESF-1-binding motif is involved in E1A 13S autoregulation of the adenovirus 12 TS2 promoter.

Pützer

Gnauck

Kirch

et al. 1997

Journal of General Virology

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Analysis of promoters of adenovirus type 12 E1A gene in a cell-free transcription system

Cited by 9 publications

References 24 publications

Direct effect of basic fibroblast growth factor on gene transcription in a cell-free system.

Direct effect of basic fibroblast growth factor on gene transcription in a cell-free system.

A novel cis-acting element that controls transcription of human immunodeficiency virus type 1 DNA, depending on cell type

A cis-acting element 7 bp upstream of the ESF-1-binding motif is involved in E1A 13S autoregulation of the adenovirus 12 TS2 promoter.

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