CLONING, SEQUENCING AND CHARACTERIZATION OF THE 5'-FLANKING REGION OFTHE HUMAN COLLAGENASE-3 GENE

Collagenase-3 (matrix metalloprotease-13) is a recently discovered hum an collagenase produced in normal articular cartilage chondrocytes and thought to be involved in the pathological process of osteoarthritis. We have sequenced and characterized 1.6 kb of the human collagenase-3 gene 5'-flanking region. The transcription start site was located 22 bp upstream from the ATG start codon. Sequence analysis of the 5'-flan king region revealed the presence of the consensus recognition sites f or the TATA and CCAAT DNA-binding proteins, activator protein-1 and E2 6 transformation specific/polyoma virus enhancer, as well as three cor e motifs of hormone response elements. Transient transfection assays d emonstrated that a small fragment of 133 bp, containing the activator protein-1 and E26 transformation specific/polyoma virus enhancer sites promoted transcription in normal and osteoarthritic human chondrocyte s with significantly higher activity than the original 1.6 kb fragment . Nucleotide sequence comparison of the promoter region of human colla genase-3 revealed a stronger similarity to the mouse collagenase-1 pro moter than to the human collagenase-1 promoter.