PMID- 1653238 OWN - NLM STAT- MEDLINE DCOM- 19911004 LR - 20210210 IS - 0021-9258 (Print) IS - 0021-9258 (Linking) VI - 266 IP - 25 DP - 1991 Sep 5 TI - Complete structure of the human gene for 92-kDa type IV collagenase. Divergent regulation of expression for the 92- and 72-kilodalton enzyme genes in HT-1080 cells. PG - 16485-90 AB - The complete structure of the human gene for 92-kDa type IV collagenase was determined. Two overlapping genomic clones spanning 26 kilobases (kb) of genomic DNA were shown to contain the entire 7.7-kb structural gene together with 15 and 3.5 kb of 5'-end and 3'-end flanking regions, respectively. The 92-kDa type IV collagenase gene contains 13 exons as does the 72-kDa type IV collagenase gene. All intron locations of the 92-kDa enzyme gene coincided with intron locations in the 72-kDa enzyme gene. Exons 5, 6, and 7 which were 174, 174, and 177 base pairs long, respectively, each encoded one complete internal repeat which resembles the collagen-binding domains of fibronectin. The sequence coding for a unique 48-residue segment in the 92-kDa type IV collagenase that has no counterpart in other metalloproteinases was not present in a separate exon, but was contained in exon 9 which also codes for sequences with homology to the other metalloproteinases. The initiation site for transcription was determined by primer extension analysis. Sequencing analysis of 599 base pairs of the 5'-end flanking region showed that the promoter does not have a TATA motif, but a TTAAA sequence at position -29 to -25. A CAAT motif was not observed but there was one GC box. Two putative 12-O-tetradecanoyl-phorbol-13-acetate (TPA) response elements, that might serve as binding sites for the transcription factor AP-1 and a consensus sequence of a transforming growth factor beta 1 (TGF-beta 1) inhibitory element were found in the promoter region. Gelatinase assay of enzyme secreted by cultured human fibrosarcoma cells (HT-1080) revealed only low levels of 92-kDa type IV collagenase activity, whereas considerable activity of the 72-kDa enzyme was present. Northern hybridization analysis confirmed these findings. Treatment of the HT-1080 cells with TPA resulted in induction of the secretion of 92-kDa type IV collagenase activity. This induction could not be significantly inhibited by concomitant incubation with TGF-beta 1. TPA and TGF-beta 1 did not markedly affect the activities of the 72-kDa enzyme. The activities of the secreted 92- and 72-kDa enzymes by HT-1080 cells correlated with the amounts of mRNA as estimated by Northern analyses. FAU - Huhtala, P AU - Huhtala P AD - Biocenter, University of Oulu, Finland. FAU - Tuuttila, A AU - Tuuttila A FAU - Chow, L T AU - Chow LT FAU - Lohi, J AU - Lohi J FAU - Keski-Oja, J AU - Keski-Oja J FAU - Tryggvason, K AU - Tryggvason K LA - eng SI - GENBANK/M68343 SI - GENBANK/M68344 SI - GENBANK/M68345 SI - GENBANK/M68346 SI - GENBANK/M68347 SI - GENBANK/M68348 SI - GENBANK/M68349 SI - GENBANK/M68350 SI - GENBANK/M68351 SI - GENBANK/M68352 SI - GENBANK/M68353 SI - GENBANK/M68354 SI - GENBANK/M68355 SI - GENBANK/M69061 SI - GENBANK/M69062 SI - GENBANK/S52685 SI - GENBANK/S52981 GR - 36200/PHS HHS/United States PT - Journal Article PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, P.H.S. PL - United States TA - J Biol Chem JT - The Journal of biological chemistry JID - 2985121R RN - 0 (RNA, Messenger) RN - 0 (Transforming Growth Factor beta) RN - 9007-49-2 (DNA) RN - EC 3.4.24.3 (Microbial Collagenase) RN - NI40JAQ945 (Tetradecanoylphorbol Acetate) SB - IM MH - Amino Acid Sequence MH - Animals MH - Base Sequence MH - Blotting, Northern MH - Cloning, Molecular MH - DNA MH - Exons MH - Fibrosarcoma MH - *Gene Expression Regulation, Enzymologic MH - Humans MH - Introns MH - Kinetics MH - Microbial Collagenase/*genetics/metabolism MH - Molecular Sequence Data MH - RNA, Messenger/metabolism MH - Rats MH - Restriction Mapping MH - Sequence Alignment MH - Tetradecanoylphorbol Acetate/pharmacology MH - Transforming Growth Factor beta/pharmacology MH - Tumor Cells, Cultured EDAT- 1991/09/05 00:00 MHDA- 1991/09/05 00:01 CRDT- 1991/09/05 00:00 PHST- 1991/09/05 00:00 [pubmed] PHST- 1991/09/05 00:01 [medline] PHST- 1991/09/05 00:00 [entrez] AID - S0021-9258(18)55326-6 [pii] PST - ppublish SO - J Biol Chem. 1991 Sep 5;266(25):16485-90.