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A 65-kDa protein mediates the positive role of heme in regulating the transcription of CYP2B1/B2 gene in rat liver

Sultana, S and Nirodi, CS and Ram, N and Prabhu, L and Padmanaban, G (1997) A 65-kDa protein mediates the positive role of heme in regulating the transcription of CYP2B1/B2 gene in rat liver. In: Journal of Biological Chemistry, 272 (14). pp. 8895-8900.

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Heme deficiency precipitated by CoCl2 administration to rats leads to a striking decrease in the inducibility of CYP2B1/B2 mRNA levels and its transcription by phenobarbitone (PB), besides decreasing the basal levels. Exogenous hemin administration counteracts the effects of CoCl2 administration. The binding of nuclear proteins to labeled positive cis-acting element (-69 to -98 nucleotides) in the near 5'-upstream region of the gene is inhibited by CoCl2 administration to saline or PR-treated rats, as assessed in gel shift assays. Administration of exogenous hemin to the animal or addition in vitro to the extracts is able to overcome the effects of CoCl2 treatment. The protein mediating this effect has been purified from CoCl2 administered nuclear extracts by heparin-agarose, positive element oligonucleotide affinity, and heme affinity column chromatography. This 65-kDa protein manifests very little binding to the positive element, but in the presence of certain other nuclear proteins, shows a strong heme-responsive binding. The purified protein binds heme. It is also able to stimulate transcription of a minigene construct of the CYP2B1/B2 gene containing -179 nucleotides of the 5'-upstream region and the I exon in a cell-free system, manifesting heme response. It is concluded that the 65-kDa protein mediates the constitutive requirement of heme for the transcription of CYP2B1/B2 gene.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to American Society for Biochemistry and Molecular Biology
Department/Centre: Division of Biological Sciences > Biochemistry
Depositing User: Mr. S Muthuraja
Date Deposited: 22 Jan 2009 06:17
Last Modified: 19 Sep 2010 04:59
URI: http://eprints.iisc.ac.in/id/eprint/17983

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