Cloning and characterization of human cathepsin L promoter.
Cathepsin L is a lysosomal cysteine protease, which is over-expressed and secreted by malignant cells. 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. The structural portion of the human cathepsin L (hCATL) gene was cloned to elucidate its genomic organization (Chauhan et al., J. Biol. Chem. 218 (1993) 1039). In the present study, a 1.90 kb DNA fragment, containing 1825 bp of the 5' upstream region of hCATL and 75 bases of the first exon of the hCATL, was amplified by PCR from an adaptor ligated placental genomic library. This fragment has been demonstrated to exhibit promoter activity by luciferase reporter assays. Sequence analysis of this fragment revealed the presence of approximately 29 different putative transcription factor binding sites. Several of them like AP-4, GATA-1, Lmo2, CEBPB, MZF-1, NF-AT, etc. were present more than once in this region. However, a consensus CAAT box but no consensus TATA box was found within the 1.0 kb upstream of exon 1. The transcription initiation site of hCATL, using placental total RNA, was mapped to a single adenine residue 289 bases upstream of the ATG codon.
Bakhshi
R
R
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi 110029, India.
Goel
A
A
Seth
P
P
Chhikara
P
P
Chauhan
S S
SS
D024506:5' Flanking Region / Q000235:genetics; D001483:Base Sequence; D056668:Cathepsin L; D002403:Cathepsins / Q000235:genetics*; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases; D004247:DNA / Q000737:chemistry / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D024363:Transcription Initiation Site; D014407:Tumor Cells, Cultured
D011993:Recombinant Fusion Proteins; D004247:DNA; D008156:Luciferases; D002403:Cathepsins; D003546:Cysteine Endopeptidases; C534281:CTSL protein, human; D056668:Cathepsin L