@entity32768 ( @entity32768 ) is a recently cloned member of the matrix metalloproteinase family (Lohi et al., J. Biol. Chem. 276 (2001) 10134). It is expressed at highest levels in the skin by basal and suprabasal keratinocytes, and in testis by developing germ cells. To characterize the @entity32768 promoter, we isolated a 3.0 kb fragment of @entity1 genomic DNA containing 5'-flanking sequence of the @entity32768 gene, and a corresponding 660 bp fragment from the @entity19 . The 5'-flanking sequences contain no typical TATA-boxes or CCAAT sequences close to the translation initiation sites. RNase protection assay revealed that two transcription start sites are utilized in the @entity1 @entity32768 gene, situated 210 and 230 bp upstream from the translation start site. The promoter contains a GT-box, situated 300 bp upstream from the translation start site, with homology to the consensus binding site for transcription factors of the Sp family. This site is perfectly conserved between the @entity1 and @entity19 promoters. For reporter gene assays a series of constructs with fragments of increasing length of the @entity32768 promoter were coupled to the firefly luciferase gene. Reporter gene assays indicated that deletion or mutation of the GT-box dramatically reduces the transcriptional activity both in keratinocytes and in spermatogonia. Gel mobility shift assays showed that several nuclear proteins bind specifically to this sequence. Supershift assays with antibodies specific for members of the Sp family identified @entity3419 and @entity28549 as components of these protein/DNA complexes and hence as possible regulators of the @entity32768 gene. Our results indicate that the @entity32768 promoter has distinctive structural and functional features, which may control the unique expression and regulation patterns of the @entity32768 gene.

Promoter characterization of the @entity1 and XXXX epilysin (MMP-28) genes.

@entity1 :: ('9606', 'Species') :: ['human'] @entity19 :: ('10090', 'Species') :: ['mouse'] @entity3419 :: ('6667', 'Gene') :: ['Sp1'] @entity28549 :: ('6670', 'Gene') :: ['Sp3'] @entity32768 :: ('79148', 'Gene') :: ['epilysin', 'MMP-28', 'Epilysin']
@entity19 :: (10090,Species) :: ['mouse']
