Regulation of Divergent Transcription of the Genes Coding for Basement Membrane Type IV Collagen

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Abstract
The genes coding for the two polypeptide chains, alpha 1(IV) and alpha 2(IV), which form together the molecule of the basement membrane type IV collagen, were found to have a special and unusual genomic arrangement. The two genes are very closely linked, they are transcribed in opposite directions, and they apparently use a common and bidirectional promoter with a length of 127 bp. This region is characterized by a symmetrical arrangement of typical elements and by the palindromic structure of the sequence. In accordance with the symmetry of the promoter itself, a symmetrical organization of sequence motifs (SP1, CCAAT) was also observed in flanking regions. For the promoter and the flanking regions we could detect specific binding of nuclear factors that indicates their involvement in transcriptional activation. This suggests that the intrinsic symmetry of the type IV collagen promoter and its flanking regions may be a structural prerequisite for its bidirectional function. In transient gene expression systems no significant activity of the type IV collagen promoter was observed in either direction. This implies that additional enhancing elements are essential for the efficient and tissue-specific transcription of both type IV collagen genes. The screening for such controlling elements within the alpha 1(IV) and the alpha 2(IV) gene led to the observation that the transcription in direction of the alpha 2(IV) gene is activated by an element located in the first intron of the alpha 2 gene. Its enhancing effect is strictly dependent on the intact genomic structure of this region. Alternation of orientation and distance to the promoter destroys its activity completely. This element, located about 100-600 bp downstream from the start site of alpha 2(IV) transcription, seems to form a synergistically acting unit with the common promoter, essential for transcriptional activity in alpha 2 direction. We have not found additional enhancing elements in other regions of both genes. Explanations for the discrepancy with previous data, which define an enhancing element within the first intron of the alpha 1(IV) gene of mouse, are only speculative at present.