Two independent pathways for transcription from the MMTV promoter

Möws, Christian C.; Preiß, Thomas; Slater, Emily P.; Cao, Xinan; Verrijzer, C. Peter; Vliet, Peter C. van der; Beato, Miguel

doi:10.1016/0960-0760(94)90111-2

Cited by 24 publications

(11 citation statements)

References 37 publications

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“…However, in cells carrying a single copy of the MMTV promoter integrated in chromosomal DNA and precisely positioned in a nucleosome, hormone induction does not lead to displacement of this nucleosome, but to a rearrangement which makes only the dyad axis of the nucleosome accessible for double-stranded DNase I cleavage and enables simultaneous binding of receptors, NF-1 and OTF1 (Fig. 3A) (231), our findings suggest that the organization in a nucleosome may be a prerequisite for simultaneous binding of all three factors, and therefore, may contribute to optimal induction (226). As hormone receptors and NF-1 (162), as well as NF-1 and OTF1, compete for binding to naked MMTV DNA (Fig.…”

Section: Biochemical Evidence For a Role Of Chromatin In Transcriptiomentioning

confidence: 99%

Interaction of Steroid Hormone Receptors with the Transcription Initiation Complex

1996

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Section: Biochemical Evidence For a Role Of Chromatin In Transcriptiomentioning

confidence: 99%

Interaction of Steroid Hormone Receptors with the Transcription Initiation Complex

1996

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“…Similarly, no effect was seen with an adenovirus major late promoter as template (data not shown). Addition of baculovirus-expressed GR generates a transcriptional induction as expected (29). The combined addition of GR and GRIP 170 fraction shows a synergistic increase in transcription (Fig.…”

Section: A Fraction Highly Enriched For Grip 170 Stimulates Grinducedmentioning

confidence: 90%

“…DNA templates were derived from p(C 2 AT) 19 and contain either the full-length MMTV promoter from Ϫ240 comprising the HREs (240/380) linked to a 380-bp-long G-less cassette or a truncated MMTV promoter from Ϫ80 lacking the HREs (80/280) fused to a 280-bp-long cassette, serving as internal control. In vitro transcription was carried out as described (29), except that the final polyethylene glycol concentration was at 1% and poly(dI-dC) was used as competitor at a final concentration of 19 ng/l.…”

Section: Methodsmentioning

confidence: 99%

“…Transcripts were treated as described (29), separated by 6.5% denaturing polyacrylamide gel electrophoresis, and visualized by autoradiography. Quantitation was made with a PhosphorImager (Molecular Dynamics), and relative transcription was determined with respect to the transcription of the control template.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, we prepared nuclear extracts from HeLa cells (23) that efficiently mediate induction by purified glucocorticoid receptor (29). Human GR was expressed in insect cells with a baculovirus expression system and purified (28).…”

Section: A Fraction Highly Enriched For Grip 170 Stimulates Grinducedmentioning

confidence: 99%

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A Fraction Enriched in a Novel Glucocorticoid Receptor-interacting Protein Stimulates Receptor-dependent Transcription in Vitro

Eggert

Möws

Tripier

et al. 1995

Journal of Biological Chemistry

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Glucocorticoids influence numerous cell functions by regulating gene activity. The glucocorticoid receptor (GR) is a ligand-activated transcription factor and, like any other transcription factor, does not modulate gene activity just by binding to DNA. Interaction with other proteins is probably required to enhance the establishment of a functional transcription initiation complex. To identify such proteins, we analyzed the in vitro interaction of the glucocorticoid receptor bound to a double glucocorticoid response element with nuclear proteins and describe here three interacting proteins with different molecular weights. One of them, which we named GRIP 170 (GR-interacting protein), was purified and microsequenced, and it turned out to be an unknown protein. When tested in a cell-free transcription assay, the fraction highly enriched for GRIP 170 does not influence basal promoter activity but does enhance GR induction.

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Up-regulation of hormone response of human papillomavirus type 16 expression and increased DNA-protein binding by consensus mutations of viral glucocorticoid response elements

et al. 1996

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Human papillomaviruses (HPVs) and steroid hormones are linked to the development of cervical cancer. Studies from our laboratory and others showed that the steroid glucocorticoid and progesterone hormones activated the expression of HPV type 16. This activation was attributed to the specific interaction of the glucocorticoid receptor (GR) with the three glucocorticoid response elements (GREs) in the HPV16 regulatory region. In the present study, we first examined the glucocorticoid response mediated through the GREs, using GRE consensus (GREc) mutations and expression assays from a heterologous basal promoter. Both single and triple HPV16 GREc constructs increased expression in the presence of the dexamethasone glucocorticoid in HeLa cervical carcinoma cells and primary baby rat kidney epithelial cells, in comparison with the triple wild-type GREs. Further, the hormone increased significantly the expression of the viral E6-E7 oncogene mRNA from intact HPV in primary human ectocervical cells in in situ hybridization assays. Three in vitro assays of DNA-protein interaction with oligonucleotides and HeLa cell extracts showed a higher binding of protein to two of the HPV16 GREcs than to the wild-type GREs. This applied especially to the GRE containing an overlapping NF1 half site, that also had a greater differential induction by dexamethasone of expression in vivo. The NF1 site was mutated in the GREc that also was bound by unique, lower-mobility complexes in electrophoretic mobility shift assays. UV-crosslinking assays confirmed the increased binding and showed binding by a 96-kDa protein, probably the GR. Our results show an important role of glucocorticoids in HPV16 expression. The direct action through the HPV16 GREs is suggested to be mediated by the hormone-activated GR in association with other factors.

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Two independent pathways for transcription from the MMTV promoter

Cited by 24 publications

References 37 publications

Interaction of Steroid Hormone Receptors with the Transcription Initiation Complex

Interaction of Steroid Hormone Receptors with the Transcription Initiation Complex

A Fraction Enriched in a Novel Glucocorticoid Receptor-interacting Protein Stimulates Receptor-dependent Transcription in Vitro

Up-regulation of hormone response of human papillomavirus type 16 expression and increased DNA-protein binding by consensus mutations of viral glucocorticoid response elements

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