Huhtala P, Chow L T, Tryggvason K
Biocenter University of Oulu, Finland.
J Biol Chem. 1990 Jul 5;265(19):11077-82.
The structure of the gene for human 70-kDa type IV collagenase (gelatinase) was determined. Three overlapping genomic clones were isolated and shown to contain 0.4 kilobase (kb) of the 5'-flanking region, the 27-kb structural gene, and 4.5 kb of the 3'-flanking region. The gene has 13 exons that vary in length from 110 to 901 base pairs (bp) and 12 introns that range from 175 to 4350 bp. Alignment of intron locations demonstrated that introns 1-4 and 8-12 of the type IV collagenase gene coincide with intron locations in the interstitial collagenase and stromelysin genes, indicating a close structural relationship of these metalloproteinase genes. Exons 5-7 are each 174 bp in size, and each codes for one complete internal repeat that resembles the collagen-binding domains of fibronectin. The transcription initiation site was determined by primer extension and S1 nuclease analyses. Analysis of the 0.4-kb 5'-flanking region of the gene showed that, in contrast to the genes of interstitial collagenease and stromelysin, there is no TATA box or 12-O-tetradecanoylphorbol-13-acetate-responsive element present in the promoter region, whereas there are two GC boxes. There is no CAAT box, but a potential binding site (CCCCAGGC) for the transcription factor AP-2 is located in the first exon.
已确定人70-kDa IV型胶原酶（明胶酶）基因的结构。分离出三个重叠的基因组克隆，显示其包含0.4千碱基（kb）的5'侧翼区、27-kb的结构基因和4.5 kb的3'侧翼区。该基因有13个外显子，长度从110到901碱基对（bp）不等，还有12个内含子，长度从175到4350 bp不等。内含子位置的比对表明，IV型胶原酶基因的内含子1-4和8-12与间质胶原酶和基质溶解素基因的内含子位置一致，表明这些金属蛋白酶基因存在密切的结构关系。外显子5-7的大小均为174 bp，每个外显子编码一个完整的内部重复序列，类似于纤连蛋白的胶原结合结构域。通过引物延伸和S1核酸酶分析确定了转录起始位点。对该基因0.4-kb的5'侧翼区的分析表明，与间质胶原酶和基质溶解素的基因不同，启动子区域不存在TATA盒或12-O-十四烷酰佛波醇-13-乙酸酯反应元件，而有两个GC盒。没有CAAT盒，但转录因子AP-2的一个潜在结合位点（CCCCAGGC）位于第一个外显子中。
