Dynamics of DNA-Protein Complex Formation in Rat Liver during Induction by Phenobarbital and Triphenyldioxane

V. O. Pustylnyak¹, L. Yu. Zacharova¹, L. F. Gulyaeva^1*, V. V. Lyakhovich¹, and N. M. Slynko²

¹Institute of Molecular Biology and Biophysics, ul. Timakova 2, Novosibirsk 630117, Russia; fax: (383-2) 323-147; E-mail: gulyaeva@soramn.ru

²Institute of Cytology and Genetics, pr. Lavrent'eva 10, Novosibirsk 630090, Russia; fax: (383-2) 331-278

^* To whom correspondence should be addressed.

Received February 9, 2004; Revision received March 12, 2004

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CYP2B gene expression in liver of rats treated with phenobarbital and triphenyldioxane at early stage of induction (40 min-18 h) was studied using electrophoretic mobility shift assay (EMSA) and RT-PCR. During first 6 h after induction, differences in the dynamics of formation of DNA-protein complexes were shown for each inducer. Later (18 h after induction), the intensity pattern of these complexes became the same for both phenobarbital and triphenyldioxane treated animals. This suggests the existence of specific signaling for each inducer only in early stages of CYP2B activation. Increase in nuclear protein (possible transcription factor) binding to Barbie-box regulatory sequence of CYP2B genes was accompanied by their increased expression. Thus, we have demonstrated for the first time that early stages of induction (40 min and 3 h after administration of phenobarbital and triphenyldioxane, respectively) are accompanied by activation of nuclear proteins that can bind to Barbie-box element of CYP2B. Although various chemical inducers cause distinct activation of such binding, this process involves activation of gene transcription.

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KEY WORDS: CYP2B genes, triphenyldioxane, Barbie-box sequence, nuclear extract proteins, electrophoretic mobility shift assay (EMSA), RT-PCR

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