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Human chromosome 7 ORF 24 (C7orf24) has been identified as a tumor-related protein, and shown to be ac-glutamyl cyclotransferase. In the cur-rent study, we characterized the promoter region of the human C7orf24 gene to explore the transcriptional regulation of the gene. | IFEBS Journal Multiple NF-Y-binding CCAAT boxes are essential for transcriptional regulation of the human C7orf24 gene a novel tumor-associated gene Yuji Ohno1 Akira Hattori1 Masamichi Ueda2 Susumu Kageyama3 Tatsuhiro Yoshiki4 and Hideaki Kakeya1 1 Department of System Chemotherapy and Molecular Sciences Division of Bioinformatics and ChemicalGenomics Graduate Schoolof PharmaceuticalSciences Kyoto University Japan 2 Institute for Virus Research Kyoto University Japan 3 Department of Urology Shiga University of MedicalScience Japan 4 Department of ClinicalOncology Kyoto PharmaceuticalUniversity Japan Keywords C7orf24 CCAAT box NF-Y tumor biomarker Correspondence A. Hattori H. Kakeya Department of System Chemotherapy and Molecular Sciences Division of Bioinformatics and Chemical Genomics Graduate School of PharmaceuticalSciences Kyoto University Sakyo Kyoto 606-8501 Japan Fax 81 75 753 4591 Tel 81 75 753 9267 81 75 753 4524 E-mail ahattori@pharm.kyoto-u.ac.jp scseigyo-hisyo@pharm.kyoto-u.ac.jp Received 28 June 2011 revised 12 August 2011 accepted 22 August 2011 doi 10.1111 j.1742-4658.2011.08314.x Human chromosome 7 ORF 24 C7orf24 has been identified as a tumor-related protein and shown to be a y-glutamyl cyclotransferase. In the current study we characterized the promoter region of the human C7orf24 gene to explore the transcriptional regulation of the gene. We revealed that the human C7orf24 promoter is a TATA-less promoter containing five CCAAT boxes aligned in a forward orientation. By performing a luciferase reporter assay with 5 -deleted and site-directed mutated constructs in HeLa MCF-7 and IMR-90 cells we found that three proximal CCAAT boxes are important for basal transcription. Electrophoretic mobility gel shift assay and chromatin immunoprecipitation assay demonstrated that NF-Y specifically bound to all three CCAAT boxes. In addition the mRNA and protein expression levels of C7orf24 were significantly reduced in HeLa cells depleted of NF-YB a subunit of
