Studies of biological function and relationship between structure and function of major-type sigma factors

主要类型西格玛因子的生物学功能及其结构与功能关系的研究

• 批准号: 09460046
• 负责人: TAKAHASHI Hideo
• 金额: $ 7.81万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09460046/
• 关键词: principal-type sigma factors; stationary phase; σィイD138ィエD1; Escherichia coli; cyanobacterium; Arabidopsis thaliana; plastid; red algae; 定常期シグマ因子; 葉緑体; ホロ酵素; rpoS; katE; sig遺伝子

1. Using rpoS-deficient (rpoS:: Tn10) strains, an rpoS-dependent katE promoter was delimited in a region different from that previously reported by others. Furthermore, in the promoter region a -10 consensus hexamer sequence was found, but ?35 sequence was not. A carboxy-terminal region responsible for the σ ィイD138ィエD1-speeific salt-tolerance was identified by in vitro transcription experiments and named CTE (carboxy-terminal element). 2. Group 2 sigma factors of cyanobacterium Synechococcus sp. JPCC7942 were shown to recognize E. coli canonical promoters as well as cyanobacterial common promoters by in vitro, experiments. A proteome analysis, of Synechocystis sp. PCC6803 was performed and it was suggested that sigD and sigE genes were involved in the induction of specific genes during nitrogen-deficient growth condition. 3. Multiple nuclear genes (sig) encoding plastid RNA polymerase sigma factors were identified from red algae, Cyanidium caladium RK-1 and higher plants including Arabidopsis thaliana, tobacco and rice. Structural and functional analyses of these genes were carried out. The overexpressed and purified gene products were shown to have sigma activity in vitro. For A. thaliana, three new sig genes, named sigD, sigE, and sigE, besides sigA, sigB and sigC, were identified and chromosomal locations of these genes were determined. Furthermore, using uid and GFP gene fusion and transformation into Arabidopsis plants, light inducibility, localization of gene products and spacial and temporal expression of these genes during plastid development were analyzed.

1.使用rpoS缺陷（rpoS：：Tn 10）菌株中，rpoS依赖的katE启动子被限定在与先前报道的不同的区域中。此外，在启动子区的一个-10的共识六聚体序列被发现，但？第35章不是通过体外转录实验，鉴定了一个与σ-D138 β-D1特异性耐盐性有关的羧基末端区域，命名为CTE（carboxy-terminal element）。2.蓝藻聚球藻JPCC 7942的第2组σ因子识别E.大肠杆菌典型启动子以及蓝藻常见启动子。通过对集胞藻PCC 6803蛋白质组的分析，发现sigD和sigE基因在氮缺乏条件下参与了特定基因的诱导。3.从红藻Cyanidium calcium RK-1和高等植物包括拟南芥、烟草和水稻中鉴定了编码质体RNA聚合酶σ因子的多个核基因（sig）。对这些基因进行了结构和功能分析。过表达和纯化的基因产物在体外显示出具有sigma活性。对于.在拟南芥中，除sigA、sigB和sigC基因外，还发现了3个新的sig基因，分别命名为sigD、sigE和sigE，并确定了它们的染色体定位。利用uid和GFP基因融合转化拟南芥，分析了它们在质体发育过程中的光诱导、基因产物的定位和时空表达。

项目成果

Y. Tozawa et al.: "Nuclear encoding of aplastid sigmafactorin rice and its tissue and light dependent expression"Nucleic Acids Res.. 15. 415-419 (1998)

Y. Tozawa 等：“质体 sigmafactorin 水稻及其组织的核编码和光依赖性表达”Nucleic Acids Res.. 15. 415-419 (1998)

A. Goto-Seki et al.: "Specificity cross talk amon group 1 and group 2 sigma factors in the cyanobacterium Synechoococous sp. PCC7942"Mol Microbiol.. 34. 473-484 (1999)

A. Goto-Seki 等人：“蓝藻聚胞球菌 PCC7942 中第 1 组和第 2 组西格玛因子的特异性串扰”Mol Microbiol.. 34. 473-484 (1999)

Tanaka, K., Y. Tozawa, N. Mochizuki, K. Shinozaki, A. Nagatani, K. Wakasa, and H. Takahashi: "Characterization of three cDNA, species encoding plastid RNA polymerase sigma factors in Arabidopsis thaliana : Evidence for the sigma factor heterogeneity in hi

Tanaka, K.、Y. Tozawa、N. Mochizuki、K. Shinozaki、A. Nagatani、K. Wakasa 和 H. Takahashi：“拟南芥中编码质体 RNA 聚合酶 σ 因子的三种 cDNA 的表征：证据

Tozawa, Y: "Nuclear encoding of a plastid sigma factor in rice and its tissde-and light-dependent expression" Nucleic Arids Res. 15. 415-419 (1998)

Tozawa，Y：“水稻中质体西格玛因子的核编码及其组织和光依赖性表达”核干旱研究。

K. Tanaka et al.: "Characterization of three cDNA species encoding plastid RNA polymerase sigma factors in Arabidopsis thaliana"FEBS Letters. 413. 309-313 (1997)

K. Tanaka 等人：“拟南芥中编码质体 RNA 聚合酶 σ 因子的三种 cDNA 物种的表征”FEBS Letters。