Structural analysis and promoter characterization of the human membrane-type matrix metalloproteinase-1 (MT1-MMP) gene

被引:113
作者
Lohi, J
Lehti, K
Valtanen, H
Parks, WC
Keski-Oja, J
机构
[1] Univ Helsinki, Dept Virol, Haartman Inst, FIN-00290 Helsinki, Finland
[2] Univ Helsinki, Dept Pathol, FIN-00290 Helsinki, Finland
[3] Washington Univ, Sch Med, Dept Pediat, St Louis, MO 63110 USA
[4] Washington Univ, Sch Med, Dept Cell Biol & Physiol, St Louis, MO 63110 USA
基金
美国国家卫生研究院; 芬兰科学院;
关键词
cell invasion; gelatinase A; Sp-1;
D O I
10.1016/S0378-1119(99)00549-1
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Membrane type-1 matrix metalloproteinase (MT1-MMP) degrades extracellular matrix components directly and indirectly by activation of other matrix metalloproteinases (MMPs). In the present study, we have isolated and characterized the human MT1-MMP gene and its promoter. The gene consists of 10 exons and nine introns spanning more than 10 kilobases (kb). The locations of two exon-intron splicing sites are distinct from the preserved positions among other known MMP genes. Primer extension and RNAse and S1 nuclease protection analyses indicated that there are four major and several minor transcription start sites. The 5'-flanking sequence of the gene contains putative regulatory elements, including one Sp-1 site and four CCAAT-boxes, whereas there is no TATA-box. The Sp-1 binding site was functional, as shown by gel shift and supershift analyses. Transfection studies with promoter constructs containing 0.1 to 7.2 kb of 5'-flanking sequence coupled to a luciferase reporter gene indicated that the promoter contains additional positive and negative regulatory sequences. Deletion of the Sp-1 binding site by site-directed mutagenesis reduced luciferase activity by about 90%, demonstrating the crucial role of this element in maintaining MT1-MMP transcription. Our findings indicate that the human MT1-MMP promoter has distinctive structural and functional features compared with other MMP genes, which may lead to a unique expression pattern and regulation during physiological and pathological processes. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:75 / 86
页数:12
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