Effects of phenylarsine oxide on expression of heme oxygenase-1 reporter constructs in transiently transfected cultures of chick embryo liver cells.

Abstract

Heme oxygenase catalyzes the first and rate-controlling step of heme catabolism. Induction of heme oxygenase-1 can be caused by numerous factors, including heme, other metalloporphyrins, transition metal ions, heat shock, ultraviolet light, phorbol esters, sodium arsenite, and phenylarsine oxide (PAO). Induction of this enzyme may protect cells from oxidative damage. Using heme oxygenase-1 promoter/reporter gene constructs, we have previously reported that the sodium arsenite-mediated induction of heme oxygenase-1 in chick embryo liver cells and chicken hepatoma (LMH) cells involves an AP-1 element. We have now investigated whether the PAO-mediated induction of heme oxygenase-1 also involves an AP-1 element. Primary cultures of chick embryo liver cells were transiently transfected with heme oxygenase-1 promoter/reporter gene constructs, treated with PAO, and reporter gene activities were measured. We found that the PAO-mediated increase in reporter gene activity was dose- and time-dependent. This activity was decreased by prior treatment with N-acetylcysteine. Studies with mutated constructs showed that both an AP-1 element and a metal responsive element are involved in the PAO-mediated induction of the heme oxygenase-1 reporter construct. Electrophoretic mobility shift assays showed that nuclear proteins from PAO-treated cells had increased binding to an AP-1 probe, and that this increase was abrogated by N-acetylcysteine. These findings support the hypothesis that the PAO-mediated induction of heme oxygenase-1 is caused by activation of AP-1 and MRE/cMyc elements and may involve nuclear proteins whose states of phosphorylation determine binding to regulatory elements, and thus the level of expression of heme oxygenase-1.

• Authors: Y Shan, R W Lambrecht, T Hong Lu, H L Bonkovsky
• Journal: Archives of biochemistry and biophysics (Arch Biochem Biophys) Vol. 372 Issue 2 Pg. 224-9 (Dec 15 1999) ISSN: 0003-9861 [Print] United States
• PMID: 10600159 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)
• Copyright: Copyright 1999 Academic Press.

Chemical References

• Arsenicals
• Nuclear Proteins
• Proto-Oncogene Proteins c-myc
• Transcription Factor AP-1
• oxophenylarsine
• DNA
• Heme Oxygenase (Decyclizing)
• Heme Oxygenase-1
• Protein Tyrosine Phosphatases
• Acetylcysteine

Topics

• Acetylcysteine (pharmacology)
• Animals
• Arsenicals (antagonists & inhibitors, pharmacology)
• Binding, Competitive
• Cells, Cultured
• Chick Embryo
• DNA (genetics, metabolism)
• Dose-Response Relationship, Drug
• Gene Expression Regulation, Enzymologic (drug effects)
• Genes, Reporter (genetics)
• Heme Oxygenase (Decyclizing) (genetics)
• Heme Oxygenase-1
• Liver (cytology, drug effects, metabolism)
• Mutation (genetics)
• Nuclear Proteins (metabolism)
• Promoter Regions, Genetic (genetics)
• Protein Tyrosine Phosphatases (antagonists & inhibitors)
• Proto-Oncogene Proteins c-myc (physiology)
• Response Elements (genetics)
• Transcription Factor AP-1 (metabolism)
• Transfection