Abstract
To elucidate the molecular basis underlying the difference in the mode of gene expression between mouse complement C4 (constitutive) and sex-limited protein (Slp) (testosterone-regulated), we compared nucleotide sequences and transcriptional regulatory activities of the 5'-flanking regions of these two genes. Although the two sequences showed a high degree of overall homology (95%) up to 1.9 kilobases (kb) upstream from the transcription initiation site, the Slp sequence lacked a 31-nucleotide segment containing ACACCC repeats and a 60-nucleotide segment containing ACAC repeats, which are present, respectively, 1.6 kb and 200 base pairs (bp) upstream from the transcription initiation site of the C4 gene. When assayed in human hepatoma-derived HepG2 cells, the 1.8-kb 5'-flanking DNA fragment of the C4 gene demonstrated strong transcriptional activity, whereas the corresponding DNA fragment of the Slp gene showed only negligible activity. By progressive deletion experiments, it was shown that the difference in the constitutive transcriptional activity of the C4 and Slp genes was accounted for by the presence or absence of the positive regulatory domain located between 1700 bp and 400 bp upstream of the transcription initiation site.