ABSTRACT
Potential matrix-associated elements in the 5'-region of the rat estrogen sulfotransferase gene (Ste1) were searched for and characterized. The DNA fragments corresponding to the regions -800/+1048 and +1049/+2038 relative to the main point of transcription initiation were found to be bound to the nuclear matrix in vitro. A permanent association of the 5'-region of the Ste1 gene with the nuclear matrix in rat hepatocytes was found, the most probable site of attachment being the region -352/-152. No differences were found in the attachment of the 5'-region of the Ste1 gene to the nuclear matrix in the liver of males where the gene is actively transcribed and in that of females where it is inactive.
Subject(s)
5' Flanking Region/genetics , Nuclear Matrix/genetics , Sulfotransferases/genetics , Animals , Chlorocebus aethiops , Female , Hepatocytes/enzymology , Male , Nuclear Matrix/enzymology , Rats , Sex Factors , Sulfotransferases/metabolismABSTRACT
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.
Subject(s)
Liver/metabolism , Nuclear Proteins/metabolism , Sulfotransferases/genetics , Animals , Base Sequence , Binding Sites/genetics , Binding, Competitive , Electrophoretic Mobility Shift Assay , Female , Male , Molecular Sequence Data , Oligonucleotides/genetics , Oligonucleotides/metabolism , Protein Binding , Rats , Sequence Homology, Nucleic Acid , Sex Factors , Sulfotransferases/metabolismABSTRACT
Estrogen sulfotransferases (STE) are members of a large superfamily of sulfotransferases. The expression of rat Ste genes is regulated in a tissue- and sex-specific manner. The cDNAs of two closely related rat estrogen sulfotransferases have been cloned earlier. By PCR performed on rat genomic DNA, we have amplified fragments of two estrogen sulfotransferase genes, Ste1 and Ste2, and established their exon-intron organization. By rapid amplification of the 5'-terminal cDNA sequences, the two variants of 5'-untranslated regions of both Ste1 and Ste2 mRNA were found. These variants are produced through transcription initiation from sites located in front of the alternative exons 1a and 1b and alternative splicing. Intron 1 of Ste1 gene contains the repeated element of ID family inserted after duplication of the ancestor gene.
