tailieunhanh - Báo cáo khoa học: Ets-1/ Elk-1 is a critical mediator of dipeptidyl-peptidase III transcription in human glioblastoma cells

Dipetidyl-petidase III is a metallopeptidase involved in a number of physi-ological processes and its expression has been reported to increase with the histological aggressiveness of human ovarian primary carcinomas. Because no information regarding the regulation of its expression was available, experiments were designed to clone, define and characterize the promoter region of the human dipeptidyl-peptidase III (DPP-III) gene. | ỊFEBS Journal Ets-1 Elk-1 is a critical mediator of dipeptidyl-peptidase III transcription in human glioblastoma cells Abhay A. Shukla Misti Jain and Shyam S. Chauhan Department of Biochemistry All India Institute of MedicalSciences New Delhi India Keywords 5 -RACE electrophoretic mobility shift assays promoter site-directed mutagenesis transcription factors Correspondence S. S. Chauhan Room No. -3009 Department of Biochemistry All India Institute of MedicalSciences New Delhi 110029 India Fax 91 11 2658 8663 Tel 91 11 2659 3272 E-mail s_s_chauhan@ Database The nucleotide sequence of the human DPP-III promoter has been submitted to the GenBank database under the accession number FJ793449 Received 9 November 2009 revised 25 December 2009 accepted 1 February 2010 doi Dipetidyl-petidase III is a metallopeptidase involved in a number of physiological processes and its expression has been reported to increase with the histological aggressiveness of human ovarian primary carcinomas. Because no information regarding the regulation of its expression was available experiments were designed to clone define and characterize the promoter region of the human dipeptidyl-peptidase III DPP-III gene. In this study we cloned a 1038 bp 5 -flanking DNA fragment of the human DPP-III gene for the first time and demonstrated strong promoter activity in this region. Deletion analysis revealed that as few as 45 nucleotides proximal to the transcription start site retained 40 of the activity of the full-length promoter. This promoter lacked the TATA box but contained multiple GC boxes and a single CAAT box. Similarly two Ets-1 Elk-1-binding motifs are present in the first 25 nucleotides from the transcription start site. Binding of Ets-1 Elk-1 proteins to these motifs was visualized by electrophoretic mobility shift and chromatin immunoprecipitation assays. Mutations of these binding sites abolished not only binding of the Ets protein but also the

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