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.

黄瓜第一叶的生长受到UV-B的阻碍。有人提出，DNA损伤的光恢复酶与植物的紫外线耐受性有关，因为迟滞的作用谱类似于光造成的DNA损伤的光谱。因此，我研究了光恢复酶（光溶酶）的基因表达的控制系统。我分离了编码CPD-光解酶的cDNA，以及来自黄瓜植物的基因组DNA（CSPHR），CSPHR的转录水平（mRNA）在光周期中的早晨达到了最大值。 uv-b添加的峰值变得更高。I使光诱导CSPHR诱导的作用光谱，并且该光谱显示了310nm左右的诱导最高峰。接下来，我将嵌合基因GUS基因作为与CSPHR启动子绑定的记者一起引入拟南芥。当将包括UV-B在内的光被照射到这些转基因植物时，GUS在叶子，茎和根部表示。在这种情况下，在GUS诱导的动作范围中，观察到了诱导量最突出的峰值。引起了携带记者Gene Luc的启动子的合并衍生物，以阐明启动子的顺式域，并检查了UV-B的LUC诱导。已经发现，翻译起点的202-296bp上流流的启动子域是诱导至关重要的顺式域。但是，还表明，仅此领域就无法诱导记者，并建议还有其他因素与翻译开始的201BP上流流相同。

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