Transcription of Cathepsin B in Glioma Cells: Regulation by an E-Box Adjacent to the Transcription Initiation Site
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| 024 | 7 | 0 | |a 10.1515/BC.2003.157 |2 doi |
| 035 | |a (NATIONALLICENCE)gruyter-10.1515/BC.2003.157 | ||
| 245 | 0 | 0 | |a Transcription of Cathepsin B in Glioma Cells: Regulation by an E-Box Adjacent to the Transcription Initiation Site |h [Elektronische Daten] |c [S. Yan, D. T. Jane, M. J. Dufresne, B. F. Sloane] |
| 520 | 3 | |a We have previously isolated the human cathepsin B promoter and shown that Sp1 and Ets factors are involved in the regulation of cathepsin B expression. Using mutagenesis, transient transfection and electrophoretic mobility shift assays (EMSAs), we further identified regulatory factors that mediate cathepsin B transcription in U87 human glioblastoma cells. An Ebox element (CACGTG) adjacent to the transcription initiation site (at nucleotides -7 to -2) was found to be indispensable for cathepsin B promoter activity. Mutation of this E-box element in both pSCB2, a promoter construct with high promoter activity, and pSCB6, a construct with basal promoter activity, led to a 90% decrease in promoter activity in U87 cells. EMSAs demonstrated that upstream stimulatory factor 1 (USF-1) and upstream stimulatory factor 2 (USF-2) bound to the Ebox as a heterodimer. Chromatin immunoprecipitation assays revealed that both USF-1 and USF-2 were associated with the cathepsin B promoter. The roles of USF-1 and USF-2 in the regulation of cathepsin B expression were demonstrated by (i) cotransfection experiments showing that USF-1 or USF-2 increased promoter activity by 2.5-fold individually and by 3.4-fold together; (ii) cotransfection of pSCB6 with pUSF-2δN (a dominant negative USF-2 expression plasmid) resulting in an 80% decrease in promoter activity; and (iii) mutation of the Ebox element (from 5'-CACGTG to 5'-CGCGTT in the pSCB6 basal promoter construct) abolishing transactivation of cathepsin B by USF-1 and USF-2. These results collectively indicate that an Ebox at nucleotides 7 to 2 of the cathepsin B promoter is critical to the expression of cathepsin B and that binding of USF-1 and USF-2 to this E-box can regulate cathepsin B promoter activity. | |
| 540 | |a Copyright © 2003 by Walter de Gruyter GmbH & Co. KG | ||
| 690 | 7 | |a Biochemistry |2 nationallicence | |
| 690 | 7 | |a Molecular biology |2 nationallicence | |
| 690 | 7 | |a Cellular biology |2 nationallicence | |
| 700 | 1 | |a Yan |D S. |4 aut | |
| 700 | 1 | |a Jane |D D. T. |4 aut | |
| 700 | 1 | |a Dufresne |D M. J. |4 aut | |
| 700 | 1 | |a Sloane |D B. F. |4 aut | |
| 773 | 0 | |t Biological Chemistry |d Walter de Gruyter |g 384/10-11(2003-11-07), 1421-1427 |x 1431-6730 |q 384:10-11<1421 |1 2003 |2 384 |o bchm | |
| 856 | 4 | 0 | |u https://doi.org/10.1515/BC.2003.157 |q text/html |z Onlinezugriff via DOI |
| 908 | |D 1 |a research article |2 jats | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1515/BC.2003.157 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Yan |D S. |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Jane |D D. T. |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Dufresne |D M. J. |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Sloane |D B. F. |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Biological Chemistry |d Walter de Gruyter |g 384/10-11(2003-11-07), 1421-1427 |x 1431-6730 |q 384:10-11<1421 |1 2003 |2 384 |o bchm | ||
| 900 | 7 | |b CC0 |u http://creativecommons.org/publicdomain/zero/1.0 |2 nationallicence | |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-gruyter | ||