TITLE

Transcription of Human Cathepsin B Is Mediated by Sp1 and Ets Family Factors in Glioma

AUTHOR(S)
Yan, Shiqing; Berquin, Isabelle M.; Troen, Bruce R.; Sloane, Bonnie F.
PUB. DATE
February 2000
SOURCE
DNA & Cell Biology;Feb2000, Vol. 19 Issue 2, p79
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Cathepsin B expression is increased at both the mRNA and protein levels in a wide variety of tumors. The mechanisms responsible for this regulation are not well elucidated. We have isolated a 2.2-kb cathepsin B genomic fragment that contains the 5' -flanking region of the cathepsin B gene. Using reporter gene analysis in human glioblastoma U87MG cells, we have mapped a 228-bp fragment (-172 to +56) having high promoter activity. This promoter region has a high G+C content; contains potential Sp1, Ets, and USF binding motifs; and lacks canonical TATA and CAAT boxes immediately upstream of the major transcriptional initiation site. Cotransfection experiments demonstrated that Sp1 and Ets1 could trans-activate cathepsin B transcription, whereas Ets2 could not. Electrophoretic mobility shift assays and supershift assays revealed that three of the four putative Sp1 sites in this promoter region form a specific complex containing the Sp1 transcription factor. Mutating all four of the Sp1 binding sites individually markedly reduced the promoter activity of transfected reporter genes in U87 cells. Cotransfection of this cathepsin B promoter construct with Sp1 family expression vectors in Schneider's Drosophila line 2 (SL2) cells demonstrated that Sp1 and Sp3, but not Sp4, activated cathepsin B transcription. Taken together, these results suggest that Sp1, Sp3, and Ets1 are important factors in cathepsin B transcription. The regulation of cathepsin B transcription by Sp1- and Sp1- related factors is mediated through multiple GC boxes.
ACCESSION #
5322776

 

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