TAILIEUCHUNG - Báo cáo khoa học: Sp3 transcription factor is crucial for transcriptional activation of the human NOX4 gene

NOX is the catalytic subunit of NADPH oxidase, the superoxide-generat-ing enzyme. Among several isoforms of NOX, NOX4 is abundantly expressed in various tissues. To clarify the mechanisms of constitutive and ubiquitous expression of NOX4, the promoter activities of the human NOX4gene were analyzed by reporter assays. | 1FEBS Journal Sp3 transcription factor is crucial for transcriptional activation of the human NOX4 gene 12 3 2 2 2 Masato Katsuyama Hideyo Hirai Kazumi Iwata Masakazu Ibi Kuniharu Matsuno Misaki Matsumoto2 and Chihiro Yabe-Nishimura2 1 Radioisotope Center Kyoto PrefecturalUniversity of Medicine Japan 2 Department of Pharmacology Kyoto PrefecturalUniversity of Medicine Japan 3 Department of Transfusion Medicine and CellTherapy Kyoto University Hospital Japan Keywords GC-box NADPH oxidase NOX4 Sp3 transcriptional regulation Correspondence M. Katsuyama Radioisotope Center Kyoto PrefecturalUniversity of Medicine Kyoto 602-8566 Japan Fax 81 75 251 5381 Tel 81 75 251 5381 E-mail mkatsuya@ Received 6 December 2010 revised 7 January 2011 accepted 11 January 2011 doi NOX is the catalytic subunit of NADPH oxidase the superoxide-generat-ing enzyme. Among several isoforms of NOX NOX4 is abundantly expressed in various tissues. To clarify the mechanisms of constitutive and ubiquitous expression of NOX4 the promoter activities of the human NOX4 gene were analyzed by reporter assays. The 5 -flanking and non-coding regions of the human NOX4 gene are known to contain multiple GC bases. Among them three GC-boxes containing putative Sp Klf-binding sites which were not found in rodent genes were suggested to be essential for the basal expression of the NOX4 gene in SH-SY5Y and HEK293 cells. Electrophoresis mobility shift assays demonstrated that Sp1 and Sp3 could bind to GC-boxes at positions -239 -227 and 69 81 in these cells. Chromatin immunoprecipitation assays showed that Sp1 and Sp3 could also bind to GC-boxes at positions -239 -227 and 69 81 in vivo. The promoter activity of the NOX4 gene was reduced in SH-SY5Y and HEK293 cells by transfection of an anti-Sp3 short hairpin RNA-expression plasmid. Taken together these results suggest that Sp3 plays a key role in the expression of NOX4 in various cell lineages in humans. Introduction

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