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Identification of a Basal Promoter for the Human M_r 72,000 Type IV Collagenase Gene and Enhanced Expression in a Highly Metastatic Cell Line

Laboratory of Pathology, National Cancer Institute, NIH, Bethesda, Maryland 20892

A basal promoter for the M_r 72,000 type IV collagenase gene was specifically defined by chloramphenicol acetyltransferase assays of a nested set of 5' upstream fragments containing the promoter region. This core promoter is TACATCT and is a noncanonical TATA box that fits the TATA consensus sequence. This sequence begins 26 base pairs in the upstream direction from the start site of transcription for the type IV collagenase gene. This basal promoter is active in the highly metastatic A2058 melanoma cell line. A putative enhancer was found between nucleotides -223 to -422 that produces a 7-fold increase in transcriptional activity in the A2058 melanoma cell line. The region immediately 5' of the basal promoter, upstream to position -422, contains a silencer and represses transcriptional activity in the nonmetastatic HT144 melanoma cell line. The results of this study are consistent with previous data that found high expression of M_r 72,000 type IV collagenase mRNA and enzymatic activity in the A2058 cell line, whereas low mRNA expression and type IV collagenase activity were found in the HT144 cell line.

¹ To whom requests for reprints should be addressed, at Molecular Hematology Branch, National Heart, Lung, and Blood Institute, Building 10, Room 7D18, Bethesda, MD 20892.

Received 7/30/91. Accepted 9/25/91.

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