A study on expression of the gene encoding photolyase for cyclobutane pyrimidine dimer and regulation mechanism of photolyase activity

环丁烷嘧啶二聚体光裂合酶基因表达及光裂合酶活性调控机制研究

• 批准号: 15570030
• 负责人: KONDO Noriaki
• 金额: $ 2.37万
• 依托单位: Teikyo University of Science & Technology (2004-2005)The University of Tokyo (2003)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15570030/
• 关键词: Action spectrum; Arabidopsis; cis element; Cucumber plant; Induction of gene expression; photolyase; Signal transduction; UV-B; 紫外線耐性; 日周変化

Growth of cucumber first leaves was retarded by UV-B. It was suggested that light recovery enzyme of DNA damage was concerned with UV tolerance of plants, because the action spectrum for growth retardation was similar to the spectrum for DNA damage caused by light. Thus, I studied to elucidate control system of gene expression of a light recovery enzyme (photolyase). I isolated the cDNA, which encoded CPD-photolyase, and the genome DNA (CsPHR) from cucumber plants The level of transcript (mRNA) of CsPHR reached a maximum in the morning in the photoperiod. The peak of this morning became higher by UV-B addition.I made the action spectrum of CsPHR induction by light, and the spectrum showed the highest peak of induction around 310nm. Next, I introduced the chimera gene, GUS gene as a reporter bound to the promoter of CsPHR, into Arabidopsis. GUS was expressed in the tip of leaves, the stem and the roots when light including UV-B was irradiated to these transgenic plants. In this case too, the most prominent peak of the induction was observed around 310nm in the action spectrum for GUS induction.Synthetic derivatives of the promoter harboring a reporter gene LUC were introduced into Arabidopsis, to clarify cis domain of the promoter, and the induction of LUC by UV-B was examined. It was found that a promoter domain of 202-296bp upper stream of the translation start point was a cis domain essential for the induction. However, it was also showed that this domain alone was not possible to induce the reporter, and it was suggested that there are other factors which work cooperatively with the domain within 201bp upper stream of the translation start.

UV-B 阻碍了黄瓜第一片叶子的生长。由于生长迟缓的作用光谱与光引起的DNA损伤的光谱相似，因此认为DNA损伤的光恢复酶与植物的紫外线耐受性有关。因此，我进行了研究以阐明光恢复酶（光裂解酶）基因表达的控制系统。我从黄瓜植物中分离出了编码CPD-光解酶的cDNA和基因组DNA（CsPHR）。CsPHR的转录物（mRNA）水平在光周期的早晨达到最大值。今天早上的峰值由于UV-B的添加而变得更高。我制作了CsPHR光诱导的作用光谱，光谱在310nm附近显示了诱导的最高峰。接下来，我将嵌合基因 GUS 基因作为与 CsPHR 启动子结合的报告基因引入拟南芥中。当包括UV-B在内的光照射到这些转基因植物时，GUS在叶尖、茎和根中表达。在这种情况下，在 GUS 诱导的作用谱中，在 310nm 附近也观察到了最突出的诱导峰。将带有报告基因 LUC 的启动子的合成衍生物引入拟南芥中，以澄清启动子的顺式结构域，并且诱导通过 UV-B 检查 LUC 的情况。发现翻译起始点上游202-296bp的启动子结构域是诱导所必需的顺式结构域。然而，也表明仅该结构域不可能诱导报告基因，并且表明还有其他因素与翻译起始点上游201bp内的结构域协同作用。

项目成果

Genomic structure of the cucumber CPD photolyase gene

黄瓜CPD光解酶基因的基因组结构

• 发表时间: 2003
• 期刊: OMICS 7・2
• 作者: M.Ioki;N.Nakajima;M.Tamaoki;S.Takahashi;N.Kondo
• 通讯作者: N.Kondo

M.Ioki, N.Nakajima, M.Tamaoki, S.Takahashi, N.Kondo: "Genomic structure of the cucumber CPD photolyase gene"OMICS. 7・2. 203-209 (2003)

M.Ioki、N.Nakajima、M.Tamaoki、S.Takahashi、N.Kondo：“黄瓜 CPD 光裂解酶基因的基因组结构” OMICS 7・2（2003）。

Wavelength-dependency of the Light-driven Transcriptional Activation of the Cucumber CPD Photolyase Gene

黄瓜 CPD 光解合酶基因光驱动转录激活的波长依赖性

• 发表时间: 2005
• 期刊: Phyton 45・4
• 作者: M.Ioki;S.Takahashi;N.Nakajima;H.Saji;K.Fujikura;M.Tamaoki;M.Aono;M.Kanna;D.Ogawa;M.Watanabe;N.Kondo
• 通讯作者: N.Kondo