Transcription of human cathepsin L mRNA species hCATL B from a novel alternative promoter in the first intron of its gene.

Cathepsin L is a lysosomal cysteine protease over-expressed in malignancy. It is very potent in degrading collagen, elastin, laminin and other components of the basement membrane and therefore, has been implicated in tumor invasion and metastasis. Two mRNA species, hCATL A and hCATL B, which contain an identical open reading frame and different 5'UTRs, were demonstrated to be encoded by the same gene located on chromosome 9q21-22. We have previously cloned and characterized the promoter responsible for the transcription of hCATL A (hCATL A promoter). However, it was not clear whether hCATL B is a splice variant of hCATL A or transcribed from a different promoter. In the present study, we demonstrate for the first time that hCATL B is transcribed from an alternate promoter (hCATL B promoter) located in the first intron of hCATL. This TATA-less promoter initiates transcription from two cytosine nucleotides present 191 and 367 bases upstream to the translation start codon. Deletion analysis revealed that the core promoter region lies upstream to these transcription initiation sites. This region contains several putative transcription factor-binding sites like AP-1, AP-4, GATA-1, Lmo2, NF-kappa B, MZF-1, NF-AT, etc. In U-87 MG cells, hCATL B promoter exhibits at least six times less activity than our previously characterized hCATL A promoter. However, this promoter is significantly more active in malignantly transformed cells as compared to its activity in untransformed cells. Thus, our results conclusively demonstrate that hCATL B mRNA is transcribed from an alternate promoter. Increased transcriptional activity from this promoter contributes to the elevated cathepsin L expression in transformed cells.