Isolation and characterization of rat cDNA clones for two distinct thyroid hormone receptors.

Murray, Michael; Zilz, N D; McCreary, N L; MacDonald, Michael J.; Towle, Howard C.

doi:10.1016/s0021-9258(18)37820-7

Cited by 217 publications

(14 citation statements)

References 42 publications

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“…Previous studies using a variety of TREs have shown that TR binds far more effectively in the presence of rat liver nuclear extract [21,25]. This enhancement is probably due to the heterodimerization of TR with RXR present in the nuclear extract, although the possibility that other proteins might be involved cannot be excluded.…”

Section: Discussionmentioning

confidence: 90%

“…The full-length cDNA of rat TR,8 [25] was ligated into the Flag-Mac expression vector, which contains the Flag recognition peptide at the N-terminus (IBI). This protein was expressed in the M15pREP E. coli strain.…”

Section: Preparatlon Of Recombinant Proteins In Escherichia Colimentioning

confidence: 99%

“…As the effects of T3 are mediated by the binding of the liganded TR to response elements, the next experiments examined the interaction of TR/3 with PEPCK-TRE. The form of the TR was used as this is the primary form expressed in the liver [25].…”

Section: Binding Of Trpl To Pepck-trementioning

confidence: 99%

“…Two complexes were formed between rat liver nuclear extract and PEPCK-TRE, one of which migrated in a similar fashion to the TR,-RXRac heterodimer. Several groups have demonstrated that nuclear extract will enhance the binding ofTR to TREs [21,25]. Therefore we tested whether rat liver nuclear extract would improve the binding of TR,?…”

Section: Binding Of Trpl To Pepck-trementioning

confidence: 99%

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Regulation of phosphoenolpyruvate carboxykinase gene transcription by thyroid hormone involves two distinct binding sites in the promoter

Park

Jerden

Bahouth

1995

Biochemical Journal

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Transcription of the gene for phosphoenolpyruvate carboxy-kinase (PEPCK) is stimulated by thyroid hormone (T3), glucagon (via cyclic AMP) and glucocorticoids. A region of the PEPCK promoter between -332 and -308 mediates the induction of transcription by T3. To characterize this region further, mutations were introduced into this region of the PEPCK promoter and the modified promoters ligated to the chloramphenicol acetyltransferase (CAT) reporter gene. Using these PEPCK-CAT vectors in transient transfections in HepG2 cells, it was found that T3 stimulates PEPCK transcription through two direct repeats of the AGGTCA motif located between nucleotides -330 and -319 [PEPCK-thyroid-hormone-responsive element (TRE)]. The beta form of the T3 receptor (TR beta) bound PEPCK-TRE as a homodimer but bound far more efficiently as a heterodimeric complex with the retinoid X receptor (RXR). An additional region called P3(I) (-250 to -234) is required for T3 responsiveness and binds members of the CCAAT-enhancer-binding protein (C/EBP) family. P3(I) contains an AGGTCA-like motif that can bind the TR beta-RXR heterodimer. Mutagenesis of this motif abolished TR beta-RXR binding without reducing T3 induction. Mutation of the C/EBP-binding site or insertion of a cyclic AMP-responsive-binding-protein site at P3(I) eliminated the T3 response. Our results indicate that T3 stimulation of PEPCK transcription is mediated by TR beta bound to PEPCK-TRE and requires C/EBP to be bound at the P3(I) site.

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Section: Discussionmentioning

confidence: 90%

Section: Preparatlon Of Recombinant Proteins In Escherichia Colimentioning

confidence: 99%

Section: Binding Of Trpl To Pepck-trementioning

confidence: 99%

Section: Binding Of Trpl To Pepck-trementioning

confidence: 99%

See 2 more Smart Citations

Regulation of phosphoenolpyruvate carboxykinase gene transcription by thyroid hormone involves two distinct binding sites in the promoter

Park

Jerden

Bahouth

1995

Biochemical Journal

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“…To generate the histidine-tagged RARα, the cDNA of the human RARα [19] was ligated into the KpnI and SmaI sites of pQE32 which introduces a histidine tag on the N-terminus (Qiagen). Synthesis of His-rTRβ was performed by digestion of the rat TRβ cDNA [20] with BamHI and PstI and ligation of this fragment into pQE30 (Qiagen). In this vector, the first 63 amino acids were removed from the N-terminus of rTRβ, but the DNAand ligand-binding domains were intact.…”

Section: Preparation Of Recombinant Proteins In Escherichia Colimentioning

confidence: 99%

CCAAT-enhancer-binding protein α (C/EBPα) is required for the thyroid hormone but not the retinoic acid induction of phosphoenolpyruvate carboxykinase (PEPCK) gene transcription

Park

Song

Olive

et al. 1997

Biochemical Journal

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Transcription of the gene for phosphoenolpyruvate carboxykinase (PEPCK) is stimulated by cAMP, the thyroid hormone tri-iodothyronine (T3) and retinoic acid (RA). Regulation of PEPCK transcription by T3 involves two sites in the promoter including a thyroid-hormone-response element (TRE) and a CCAAT-enhancer-binding protein (C/EBP) binding site called P3I. Mutation of either the TRE or P3I eliminates the T3 response. In this study, we examined the role of C/EBPs in the induction of PEPCK transcription by T3 and RA. PEPCK-CAT vectors were transfected into HepG2 cells. Co-transfection of a dominant negative C/EBP eliminated the T3 stimulation indicating that a member of the C/EBP family is required. To determine which C/EBP isoform was required, Gal4 fusion proteins were created that contained the Gal4 DNA-binding domain ligated to the transcriptional activation domain of C/EBP alpha, C/EBP beta or the cAMP-responsive-element-binding protein. A Gal4 DNA-binding site was introduced into the P3(I) site of the PEPCK-CAT vector. Only co-transfection of the Gal4-C/EBP alpha vector was able to restore T3 responsiveness to the PEPCK-CAT vector. The T3 and RA receptors are members of the nuclear receptor superfamily and bind to repeats of the AGGTCA motif. We found that the RA receptor can bind to sequences within the PEPCK-TRE and contribute to RA responsiveness of the PEPCK gene. However, the RA induction of PEPCK transcription was found to be independent of C/EBPs, further demonstrating the specificity of the involvement of C/EBP alpha in the T3 effect.

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Multiple glucocorticoid receptor transcripts in membrane glucocorticoid receptor-enriched S-49 mouse lymphoma cells

1999

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A cDNA library from plasma membrane glucocorticoid receptor-enriched (mGR(++)) S- 49 mouse T lymphoma cells was screened with full-length rat intracellular GR (iGR) cDNA, BUGR-2 antibody, and PCR amplimers to portions of the mouse GR cDNA. One or two single-base substitutions resulting in amino acid changes (which do not incapacitate the receptor) were found in all but one clone: Val437 --> Gly (located in the first zinc finger), and Glu546 --> Gly (in the steroid-binding domain). Two previously unidentified exon 1 variants (1D and 1E), and two of three previously reported variants (1A, 1B) were found to be spliced onto the common exon 2. Exon 1D- and 1E-containing transcripts were confirmed by direct sequencing of amplimers from reverse transcriptase-coupled PCR. RNase protection studies revealed that one of these transcripts was expressed in mGR(++) cells only, but not in two mGR-less (mGR(--) S-49, and AtT-20 mouse pituitary) cell lines. These studies suggest that at least four promoters may be responsible for the control of GR (iGR and mGR) types in mouse lymphoma cells.

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Isolation and characterization of rat cDNA clones for two distinct thyroid hormone receptors.

Cited by 217 publications

References 42 publications

Regulation of phosphoenolpyruvate carboxykinase gene transcription by thyroid hormone involves two distinct binding sites in the promoter

Regulation of phosphoenolpyruvate carboxykinase gene transcription by thyroid hormone involves two distinct binding sites in the promoter

CCAAT-enhancer-binding protein α (C/EBPα) is required for the thyroid hormone but not the retinoic acid induction of phosphoenolpyruvate carboxykinase (PEPCK) gene transcription

Multiple glucocorticoid receptor transcripts in membrane glucocorticoid receptor-enriched S-49 mouse lymphoma cells

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