Sensitivity of virally-driven luciferase reporter plasmids to members of the steroid/thyroid/retinoid family of nuclear receptors

Lynn M. Everett, David Crabb

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

During a series of transfection experiments, the pRSV-luc plasmid used as an internal control was found to be sensitive to co-transfection with expression vectors for several members of the steroid/thyroid/retinoid superfamily of nuclear receptors. Therefore, a survey of the effect of these expression vectors on the activity of four reporter plasmids was conducted. In CV-1 cells, the activity of pRSV-luc, which contains the P. pyralis luciferase gene, was repressed by co-transfection of PPARα and ARP-1 and was activated by COUP-TFI. Expression of pSV40-luc, containing the same luciferase gene, was repressed by PPARα and HNF-4 and activated by both COUP-TFI and ARP-1. All four of these expression vectors reduced the expression of the pRL-TK plasmid, which contains the luciferase gene from Renilla reniformis. RXR expression vectors had no effect on luciferase activity in CV-1 cells but induced luciferase activity in H4IIEC3 hepatoma cells. This activation was blocked by the addition of ligand, 9-cis retinoic acid. pSV2-CAT, which contains the chloramphenicol acetyltransferase gene, was insensitive to all receptor expression vectors tested. Both the P. pyralis and R. reniformis luciferase genes appear to contain sequences that render them responsive to steroid/thyroid/retinoid nuclear receptors.

Original languageEnglish
Pages (from-to)197-201
Number of pages5
JournalJournal of Steroid Biochemistry and Molecular Biology
Volume70
Issue number4-6
DOIs
StatePublished - Sep 1999

Fingerprint

Retinoids
Cytoplasmic and Nuclear Receptors
Luciferases
Thyroid Gland
Plasmids
Steroids
Genes
Transfection
Peroxisome Proliferator-Activated Receptors
Renilla Luciferases
Chloramphenicol O-Acetyltransferase
Hepatocellular Carcinoma
Ligands
Chemical activation
Experiments
1,2-diamino-1,2-N,N'-carbonyl-1,2-dideoxyglucose hydrate

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology

Cite this

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abstract = "During a series of transfection experiments, the pRSV-luc plasmid used as an internal control was found to be sensitive to co-transfection with expression vectors for several members of the steroid/thyroid/retinoid superfamily of nuclear receptors. Therefore, a survey of the effect of these expression vectors on the activity of four reporter plasmids was conducted. In CV-1 cells, the activity of pRSV-luc, which contains the P. pyralis luciferase gene, was repressed by co-transfection of PPARα and ARP-1 and was activated by COUP-TFI. Expression of pSV40-luc, containing the same luciferase gene, was repressed by PPARα and HNF-4 and activated by both COUP-TFI and ARP-1. All four of these expression vectors reduced the expression of the pRL-TK plasmid, which contains the luciferase gene from Renilla reniformis. RXR expression vectors had no effect on luciferase activity in CV-1 cells but induced luciferase activity in H4IIEC3 hepatoma cells. This activation was blocked by the addition of ligand, 9-cis retinoic acid. pSV2-CAT, which contains the chloramphenicol acetyltransferase gene, was insensitive to all receptor expression vectors tested. Both the P. pyralis and R. reniformis luciferase genes appear to contain sequences that render them responsive to steroid/thyroid/retinoid nuclear receptors.",
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