Bacterial thermosensor protein : studies on the functions of HrcA protein as a transcriptional regulator for chaperone genes.

细菌热敏蛋白：研究 HrcA 蛋白作为伴侣基因转录调节因子的功能。

• 批准号: 13660097
• 负责人: WATANABE Kunihiko
• 金额: $ 2.05万
• 依托单位: Kyoto Prefectural University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660097/
• 关键词: HrcA; CIRCE; thermosensor; DnaK; GroEL; 熱センター

In the heat-shock response of bacillary cells, HrcA repressor proteins negatively control the expression of the major heat-shock genes, groE and dnaK operons by binding the CIRCE (controlling inverted repeat of chaperone expression) element. The studies on two critical but yet unrevealed issues related to the structure and function of HrcA were performed mainly using the HrcA from obligate thermophile Bacillus thermoglucosidasius KP1006. These two critical issues are : (i) the identification of the region for HrcA to bind the CIRCE element, and (ii) whether HrcA can play the role of a thermosensor. We assigned the position of a helix-turn-helix (HTH) motif in B. thermoglucosidasius HrcA, which is one of the typical for DNA-binding proteins, and indicated that two residues in the HTH motif are crucial for the binding of HrcA to the CIRCE element. Furthermore, we compared the thermostability of the HrcA-CIRCE complex derived from Bacillus subtilis and B. thermoglucosidasius, which grow in vastly different ranges of temperature. The profiles of their HrcA-CIRCE complexes in thermostability were quite consistent with the difference in the growth temperature of B. thermoglucosidasius and B. subtilis and, thus, suggested that the HrcA can function as a thermosensor to sense temperature change in cells.

在细菌热休克反应中，HrcA阻遏蛋白通过与CIRCE（controlling inverted repeat of chaperone expression）元件结合，负调控热休克基因groE和dnaK操纵子的表达。本文主要利用嗜热芽孢杆菌（Bacillusthermoglucosidasius）KP 1006的HrcA对HrcA的结构和功能进行了研究。这两个关键问题是：（i）HrcA与CIRCE元件结合的区域的识别，以及（ii）HrcA是否可以发挥热传感器的作用。我们指定了B中螺旋-转角-螺旋（HTH）基序的位置。Thermoglucosidasius HrcA，这是一个典型的DNA结合蛋白，并表明在HTH基序的两个残基是至关重要的HrcA的结合到CIRCE元件。此外，我们还比较了枯草芽孢杆菌和B菌的HrcA-CIRCE复合物的热稳定性。thermoglucosidasius，生长在不同的温度范围。它们的HrcA-CIRCE复合物的热稳定性曲线与B的生长温度差异相当一致。热葡萄糖苷酶和B.因此，HrcA可以作为温度传感器来感知细胞中的温度变化。

项目成果

Masafumi Hitomi, Hiroshi Nishimura, Yoshiyuki Tsujimoto, Hiroshi Matsui, and Kunihiko Watabe.: "Identification of a helix-turn-helix motif of Bacillus thermoglucosidasius HrcA essential for binding to the CIRCE element and thermostability of the HrcA-CIRC

Masafumi Hitomi、Hiroshi Nishimura、Yoshiyuki Tsujimoto、Hiroshi Matsui 和 Kunihiko Watabe。：“热葡萄糖芽孢杆菌 HrcA 的螺旋-转角-螺旋基序的鉴定对于结合 CIRCE 元件和 HrcA-CIRC 的热稳定性至关重要

Kunihiko Watanabe et al.: "Renaturation of Bacillus thermoglucosidasius HrcA repressor by DNA and thermostability of the HrcA-DNA complex in vitro"Journal of Bacteriology. 183. 155-161 (2001)

Kunihiko Watanabe 等人：“通过 DNA 实现热葡糖苷芽孢杆菌 HrcA 阻遏物的复性以及体外 HrcA-DNA 复合物的热稳定性”《细菌学杂志》。

M.Okamoto et al.: "A thermostable collagenolytic protease with a very large molecular mass produced by thermophilic Bacillus sp. strain MO-1"Applied Microbiology and Biotechnology. 57. 103-108 (2001)

M.Okamoto 等人：“由嗜热芽孢杆菌菌株 MO-1 产生的具有非常大分子量的热稳定性胶原蛋白水解蛋白酶”应用微生物学和生物技术。

Kunihiko Watanabe, Takeshi Yamamoto and Yuzuru Suzuki.: "Renaturation of Bacillus thermoglucosidasius HrcA repressor by DNA and thermostability of HrcA-DNA complex in vitro"Journal of Bacteriology. 183. 155-161 (2001)

Kunihiko Watanabe、Takeshi Yamamoto 和 Yuzuru Suzuki.：“DNA 复性热葡萄糖苷酸芽孢杆菌 HrcA 阻遏物以及体外 HrcA-DNA 复合物的热稳定性”细菌学杂志。

Kunihiko Watanabe, Kazue Miyake and Yuzuru Suzuki.: "Identification of catalytic and substrate-binding site residues in Bacillus cereus ATCC7064 oligo-1,6-glucosidase"Biosci. Biotechnol. Biochem.. 65. 2058-2064 (2001)

Kunihiko Watanabe、Kazue Miyake 和 Yuzuru Suzuki.：“蜡样芽孢杆菌 ATCC7064 寡-1,6-葡萄糖苷酶中催化和底物结合位点残基的鉴定”Biosci。