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The Radiation responsive promoter of the ${it Deinococcus radiodurans pprA}$ gene

デイノコッカス・ラジオデュランス${it pprA}$遺伝子の放射線応答性プロモーター

大庭 寛史*; 佐藤 勝也*; 柳沢 忠*; 鳴海 一成

Oba, Hirofumi*; Sato, Katsuya*; Yanagisawa, Tadashi*; Narumi, Issei

デイノコッカス・ラジオデュランス${it pprA}$遺伝子の転写開始点が、翻訳開始点上流-156, -154, -22であることを同定した。全ての転写産物の量が放射線照射で増加したことから、少なくとも2つの放射線応答性プロモーターの存在が示唆された。ルシフェラーゼレポーターアッセイにより、遠位プロモーターは翻訳開始点上流-208から-156の間に、近位プロモーターは-57から-22の位置にあることが明らかとなった。-57から-38の領域は、近位プロモーター活性に必須であり、また、-33の位置のチミンが放射線応答に重要な塩基であることがわかった。さらに、${it pprI}$遺伝子産物による${it pprA}$遺伝子の発現制御がプロモーターレベルで起こることが示唆された。

Three transcriptional start points for the ${it Deinococcus radioduans pprA}$ were located at positions -156, -154 and -22 upstream from the ${it pprA}$ translation initiation site. The amount of the three extended products increased in cells exposed to 2-kGy followed by a 0.5-h post-incubation, suggesting the existence of at least two radiation responsive promoters for ${it pprA}$ expression. A luciferase reporter assay revealed that the distal promoter is located between positions -208 and -156 from the translation initiation site, while the proximal promoter is located between positions -57 and -22. The region located between positions -57 and -38 was indispensable for proximal promoter activity. Site-directed mutagenesis of a thymine positioned at -33 resulted in severe impairment of promoter activity, and suggested that the thymine functions as a master base for the proximal radiation responsive promoter. The results also suggested that up-regulation of ${it pprA}$ expression by the ${it pprI}$ gene product is triggered at the promoter level.

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パーセンタイル:53.96

分野:Genetics & Heredity

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