Characterization of the promoter of the gene encoding human tissue inhibitor of metalloproteinases-2 (TIMP-2).
Tissue inhibitors of metalloproteinases (TIMPs) are multifunctional proteins that control the proteolytic activity of matrix metalloproteinases (MMPs). We report here the cloning and characterization of a 2.5-kb genomic fragment of the human timp-2 gene that includes 519 bp of the 5' flanking region, the first coding exon (432-bp) and part of the first intron. The 5' flanking region has several features of housekeeping genes. It has a high G-C content and is included in a typical CpG island. It also contains a TATA-like element (AATAAAA) located 23 to 37-bp upstream from a cluster of transcription start points (tsp), several Sp1 and one AP-2 motifs, and an AP-1 consensus sequence located at position -590 to -583 from the start codon. When inserted upstream from a promoterless luciferase-encoding gene, a 715-bp fragment of this 5'-flanking sequence behaved as a promoter in transiently transfected NIH3T3 and Rat-1 fibroblasts. The effect of deletions of the promoter suggested the presence of a negative control element located between positions -661 and -575. This element includes the AP-1 consensus sequence. However, treatment with phorbol did not change activity in transfected cells and did not change the timp-2 mRNA content of human HT1080 fibrosarcoma cells. A comparison with the promoter of murine timp-1 revealed several differences consistent with the fact that timp-1 and timp-2 are differentially regulated.
De Clerck
Y A
YA
Division of Hematology-Oncology, Childrens Hospital Los Angeles, CA 90054-0700.
Darville
M I
MI
Eeckhout
Y
Y
Rousseau
G G
GG
D016475:3T3 Cells / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D003062:Codon / Q000235:genetics; D005091:Exons / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors*; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics*; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D019716:Tissue Inhibitor of Metalloproteinase-2; D014162:Transfection
D003062:Codon; D011506:Proteins; D012333:RNA, Messenger; D019716:Tissue Inhibitor of Metalloproteinase-2; D008666:Metalloendopeptidases; D013755:Tetradecanoylphorbol Acetate