Structural analysis and promoter characterization of the human collagenase-3 gene (MMP13)

Journal Article · Sat Mar 01 04:00:00 EST 1997 · Genomics

DOI:https://doi.org/10.1006/geno.1996.4554· OSTI ID:518468

Pendas, A M; Balbin, M; Llano, E ^[1]

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1. Universidad de Oviedo (Spain); and others

Human collagenase-3 (MMP13) is a recently identified member of the matrix metalloproteinase (MMP) family that is expressed in breast carcinomas and in articular cartilage from arthritic patients. In this work we have isolated and characterized genomic clones coding for human collagenase-3. This gene is composed of 10 exons and 9 introns and spans over 12.5 kb. The overall organization of the collagenase-3 gene is similar to that of other MMP genes clustered at chromosome 11q22, including fibroblast collagenase (MMP-1), matrilysin (MMP-7), and macrophage metalloelastase (MMP-12), but is more distantly related to genes coding for stromelysin-3 (MMP-11), gelatinase-A (MMP-2), and gelatinase-B (MMP-9), which map outside of this gene cluster. Nucleotide sequence analysis of about 1 kb of the 5{prime}-flanking region of the collagenase-3 gene revealed the presence of a TATA box, an AP-1 motif, a PEA-3 consensus sequence, an osteoblast specific element (OSE-2), and a TGF-{beta} inhibitory element. Transient transfection experiments in HeLa and COS-1 cells with chloramphenicol acetyltransferase (CAT)-containing constructs showed that the AP-1 site is functional and responsible for the observed inducibility of the reporter gene by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). However, and in contrast to other MMP genes, no significative synergistic effect on CAT activity between the AP-1 and PEA-3 elements found in the collagenase-3 gene promoter was found. DNA binding analysis with nuclear extracts from HeLa cells revealed the formation of specific complexes between collagenase-3 promoter sequences containing the AP-1 site and nuclear proteins. The presence of this AP-1 functional site, which is able to confer responsiveness to a variety of tumor promoters and oncogene products, may contribute to explaining the high-level expression of collagenase-3 in breast carcinomas and degenerative joint diseases. 48 refs., 5 figs., 2 tabs.

• Sponsoring Organization: USDOE
• OSTI ID: 518468
• Journal Information: Genomics, Journal Name: Genomics Journal Issue: 2 Vol. 40; ISSN 0888-7543; ISSN GNMCEP
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
CARCINOMAS
CHIMERAS
COLLAGEN
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
ELECTROPHORESIS
EXONS
GENE REGULATION
GENETIC MAPPING
HEREDITARY DISEASES
HUMAN CHROMOSOMES
INTRONS
MAMMARY GLANDS
METALLOPROTEINS
METHYLATION
NUCLEOTIDES
ONCOGENES
PATIENTS
POLYMERASE CHAIN REACTION
RHEUMATIC DISEASES
STRUCTURE-ACTIVITY RELATIONSHIPS
TRANSCRIPTION
TUMOR PROMOTERS