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Journal of Biochemistry Advance Access published online on September 7, 2006
Journal of Biochemistry, doi:10.1093/jb/mvj186
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© 2006 The Japanese Biochemical Society.
Received July 26, 2006
Accepted August 31, 2006

Regular Paper

Identification of the Positive Regulatory and Distinct Core Regions of Promoters, and Transcriptional Regulation in Three Types of Mouse Phospholipid Hydroperoxide Glutathione Peroxidase

Hirotaka Imai 1 *, Makoto Saito 1, Nozomu Kirai 1, Junya Hasegawa 1, Kumiko Konishi 1, Hiroyuki Hattori 1, Masuhiro Nishimura 2, Shinsaku Naito 2, and Yasuhito Nakagawa 1

1 School of Pharmaceutical Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan
2 Division of Pharmacology, Drug Safety and Metabolism, Otsuka Pharmaceutical Factory, Inc., Naruto, Tokushima 772-8601, Japan

* To whom correspondence should be addressed.
Hirotaka Imai, E-mail: imaih{at}pharm.kitasato-u.ac.jp


  Abstract

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is transcribed into three types of mRNA, mitochondrial, non-mitochondrial and nucleolar types, from one gene by alternative transcription using different first exons, Ia and Ib. We investigated the regulatory mechanisms of the expressions of the three types of PHGPx using promoter analysis with luciferase as the reporter gene and electrophoretic mobility shift analysis. Here we report a draft of the positive regulatory region and the core promoter regions of PHGPx in several cell lines. From promoter deletion analysis we identified the three distinct core regions of mitochondrial PHGPx, non-mitochondrial PHGPx and nucleolar PHGPx. The core promoter activity of non-mitochondrial PHGPx was high in L929 cells, but relatively low for mitochondrial and nucleolar PHGPx. We also identified the positive regulatory region of mitochondrial PHGPx by deletion and mutation analysis of 5'-flanking regions of mitochondrial PHGPx. This region could regulate the promoter activity of non-mitochondrial PHGPx; however, up-regulation by this region was normally suppressed by the upstream region in somatic cells. Electrophoretic mobility shift assay demonstrated that a specific transcription factor complex bound to this region in adult testis, but not in young testis and different sizes of complexes bound to this region between testis and brain.

Keywords: Promoter analysis; positive regulatory region; selenium; glutathione peroxidase; tissue regulation.
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