Hormonal regulation of transcription of rDNA: use of nucleoside thiotriphosphates to measure initiation in isolated nuclei

Zhang, Zhen Yong; Thompson, E. Aubrey; Stallcup, Michael R.

doi:10.1093/nar/12.21.8115

Cited by 13 publications

(12 citation statements)

References 15 publications

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“…Analysis of transcription in vitro indicates that glucocorticoids regulate rRNA synthesis at the level of the initiation complex rather than by influencing the rate of elongation. We suppose that a similar situation prevails in vivo and such data as are available indicate that initiation of synthesis of pre-rRNA in isolated nuclei conforms to this expectation (18).…”

Section: Discussionsupporting

confidence: 64%

Hormonal regulation of transcription of rDNA: glucocorticoid effects upon initiation and elongationin vitro

Cavanaugh

Thompson

1985

Nucl Acids Res

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Various parameters of transcription of cloned mouse rDNA have been examined using extracts from control P1798 cells and from cells treated 24 h with 0.1 microM dexamethasone. Highly purified RNA polymerase I from either source catalyzes nucleotidyl transfer (elongation) at a rate of approximately 30 nucleotides/sec. Extracts from hormone-treated cells are capable of forming stable, preinitiation complexes. The rates of stable complex formation are the same in extracts from control and hormone-treated cells. Nevertheless, initiation of transcription does not occur in extracts from hormone-treated cells. Initiation in such extracts may be restored by the addition of a partially purified RNA polymerase I initiation factor, designated TFIC. The data indicate that initiation by RNA polymerase I is a multi-step process. The first step involves the formation of stable, preinitiation complexes, as demonstrated by a number of groups. Initiation, per se, requires an additional protein, TFIC. Glucocorticoids and perhaps other mitogenic agents regulate transcription of rDNA by influencing the amount or activity of TFIC.

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Section: Discussionsupporting

confidence: 64%

Hormonal regulation of transcription of rDNA: glucocorticoid effects upon initiation and elongationin vitro

Cavanaugh

Thompson

1985

Nucl Acids Res

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“…In situ transcription extension (run-on) assays were performed as described by Diamond and Goodman (17) and Zhang et al (18), with the following modifications. Cells were grown in T-150 flasks to a density of approximately 1.3 x 10 5 cells/cm 2 , and treated for 1 h with the indicated amounts of hormone and antagonist.…”

Section: Transcription Extension Assaysmentioning

confidence: 99%

Mechanism of Dexamethasone 21-Mesylate Antiglucocorticoid Action: II. Receptor-Antiglucocorticoid Complexes do not Interact Productively with Mouse Mammary Tumor Virus Long Terminal Repeat Chromatin

Richard-Foy¹,

Sistare²,

Riegel³

et al. 1987

Molecular Endocrinology

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We have studied the interaction of covalent dexamethasone 21-mesylate (DM) labeled, activated glucocorticoid receptor with mouse mammary tumor virus (MMTV) chromatin. Studies were performed on a murine cell line (904.13) which contains 200 copies per cell of a MMTV long terminal repeat (LTR) v-rasH casette mobilized on bovine papilloma virus based episomes. DM binds covalently to glucocorticoid receptors and displays almost full antagonist activity in this cell line. In situ transcription extension assays indicate that activated receptor-DM complex cannot stimulate LTR-initiated transcription. The receptor-DM complex also fails to induce DNase I hypersensitivity (HSR) and transcription factor loading at the MMTV promoter. Transcription activation, HSR formation, and factor binding induced with the agonist dexamethasone are blocked by covalent occupation of the receptor by DM. Although DM-activated receptor binds specifically to receptor sites on purified LTR DNA, the antagonist receptor complex does not interact productively with MMTV LTR chromatin in vivo.

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“…Transcripts generated using the thionucleoside triphosphates were purified from the reaction mixture and fractionated through a mercury-Sepharose column (Hg-Sepharose) as described by Zhang et al (23). Bound transcripts were eluted (El) with TNES buffer (0.1 M Tris-hydrochloride, pH = 8.0, 1 M NaCl, 10 mM EDTA and 1% SDS).…”

Section: (D) Transcriptions In Vitromentioning

confidence: 99%

Analysis and comparison of the internal promoter, pE, of the ilvGMEDA operons fromEscherichia coliK-12 andSalmonella typhimurium

Lopes¹,

Lawther²

1986

Nucl Acids Res

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It was previously determined that the distal portion of the ilvGMEDA operon was expressed despite the insertion of transposons into ilvG and ilvE. This observation suggested the existence of internal promoters upstream of ilvE (pE) and ilvD (pD). The internal promoter pE, responsible for part of ilvEDA expression, has been analyzed both in vivo and in vitro. Our results indicate that: pE exists in both E. coli K-12 and S. typhimurium; pE is located in the distal end of the ilvM coding sequence; the pE sequence is highly conserved in the two bacteria; the amino acid sequence of the ilvM gene product is 93% homologous between the two bacteria; transcription from pE can be demonstrated both in vivo and in vitro; the efficiency of pE is essentially equivalent in the two bacteria.

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Hormonal regulation of transcription of rDNA: use of nucleoside thiotriphosphates to measure initiation in isolated nuclei

Cited by 13 publications

References 15 publications

Hormonal regulation of transcription of rDNA: glucocorticoid effects upon initiation and elongationin vitro

Hormonal regulation of transcription of rDNA: glucocorticoid effects upon initiation and elongationin vitro

Mechanism of Dexamethasone 21-Mesylate Antiglucocorticoid Action: II. Receptor-Antiglucocorticoid Complexes do not Interact Productively with Mouse Mammary Tumor Virus Long Terminal Repeat Chromatin

Analysis and comparison of the internal promoter, pE, of the ilvGMEDA operons fromEscherichia coliK-12 andSalmonella typhimurium

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