Localization of a Site for Interaction with Hepatic Male-Specific Proteins in Two Rat Estrogen Sulfotransferase Genes

I. I. Astapova¹, T. A. Shchelkunova², I. A. Morozov¹, A. N. Smirnov^2*, and P. M. Rubtsov¹

¹Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, Moscow 119991, Russia; fax: (095) 135-1405; E-mail: rubtsov@genome.eimb.relarn.ru

²School of Biology, Lomonosov Moscow State University, Moscow 119992, Russia; fax: (095) 939-4309; E-mail: smirnov_an@hotmail.com

^* To whom correspondence should be addressed.

Received February 4, 2002; Revision received February 22, 2002

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Using electromobility shift assay the interaction of fragments of two paralogous rat estrogen sulfotransferase (Ste) genes with proteins of nuclear extracts from male and female rat liver was studied. Male-specific DNA-protein complexes were revealed with labeled oligonucleotides corresponding to fragments +1150/+1449, +1358/+1449, +1397/+1449, and +1417/+1449 of intron 1 of the Ste1 gene. The removal of a 20 bp region corresponding to the sequence +1430/+1449, or even either 5´- or 3´-terminal 5 bp of this region abolished the selective interaction of the oligonucleotides with the male-specific protein(s). According to the results of the experiments on mutual competition of the oligonucleotides, the fragment of the Ste2 gene corresponding to the sequence +1397/+1449 of the Ste1 gene formed complexes with the same male-specific protein(s) as the fragment of the Ste1 gene did. The data suggest the mapped element to participate in gender differentiation of the expression of the Ste1 and Ste2 genes.

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KEY WORDS: estrogen sulfotransferase, sex differentiation, gene expression, regulatory elements, DNA-protein interaction, liver

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