Functional analysis of Legionella Dot/Icm T4SS component DotA.
军团菌 Dot/Icm T4SS 组件 DotA 的功能分析。
基本信息
- 批准号:18590420
- 负责人:
- 金额:$ 2.57万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2006
- 资助国家:日本
- 起止时间:2006 至 2007
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Protein secretion plays a central role in bacterial pathogenesis. Legionella pneumophlia, the causative agent of Legionnaires' disease, translocated a large array of effector proteins via the Dot/Icm type IV secretion system (T4SS). Dot/Icm T4SS is composed of>20 proteins and essential for Legionella infection. However, molecular basis of type IV secretion remains largely unknown. We had demonstrated that vast majority of DotA, a Dot/Icm T4SS component, was secreted into extracellular milieu via the Dot/Icm T4SS. In bacteria, DotA is integrated into inner membrane with seven trans-membrane regions. To address how such integral inner membrane protein is secreted by T4SS, we isolated DotA point mutants defective either in DotA secretion or in DotA function. DotA function was assessed by the ability of Legionella to grow within environmental model host Acanthamoeba castellanii We established a high-throughput screening system and isolated 57 DotA mutants. All of these mutants are defective both in DotA secretion and in DotA function, suggesting that DotA secretion is indispensable for DotA function. Fine mapping of these mutants unveiled 22 independent point mutations. No specific region of DotA rich of mutations was found. Notably, these point mutations were mapped both at cytoplasmic and periplasmic regions of DotA, implying that DotA secretion involve multi-step processes. Additonally, we found that DotF, a putative core component of Dot/Icm T4SS, is dispensable for DotA secretion. DotA secretion from DotF KO strain was reduced by 10-fold. Because DotF is thought to be a core component of Dot/Icm T4SS, we examined effector translocation from DotF KO strain. The data indicated that Dot F is dispensable both for DotA secretion and for effector translocation.
蛋白质分泌在细菌发病机制中起核心作用。嗜肺军团菌是军团病的病原体,通过Dot/Icm IV型分泌系统(T4SS)使大量效应蛋白易位。Dot/Icm T4SS由bbb20蛋白组成,是军团菌感染所必需的。然而,IV型分泌的分子基础在很大程度上仍然未知。我们已经证明,绝大多数DotA (Dot/Icm T4SS成分)通过Dot/Icm T4SS分泌到细胞外环境。在细菌中,DotA被整合到具有7个跨膜区域的内膜中。为了研究这种完整的内膜蛋白是如何由T4SS分泌的,我们分离了DotA分泌缺陷或DotA功能缺陷的DotA点突变体。通过军团菌在环境模型宿主castellanacanthamoeba castellanii内的生长能力来评估其DotA功能。这些突变体在DotA分泌和DotA功能上都存在缺陷,表明DotA分泌是DotA功能不可缺少的。这些突变体的精细图谱揭示了22个独立的点突变。没有发现特异的DotA区域富含突变。值得注意的是,这些点突变在DotA的细胞质和质周区域都被定位,这意味着DotA的分泌涉及多步骤过程。此外,我们发现DotF是Dot/Icm T4SS的核心成分,对于DotA的分泌是必不可少的。DotF KO菌株的DotA分泌量减少了10倍。由于DotF被认为是Dot/Icm T4SS的核心成分,我们研究了DotF KO菌株的效应物易位。数据表明,Dot F对于DotA分泌和效应体转运都是不可缺少的。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
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Screening of novel protein substrates of the Legionella Dot/Icm type IV secretion system based on the properties of C-terminal translocation signals.
基于C端易位信号的特性筛选军团菌Dot/Icm IV型分泌系统的新型蛋白质底物。
- DOI:
- 发表时间:2006
- 期刊:
- 影响因子:0
- 作者:Mun H-S.;Aosai F;Fang H;Piao LX;Winn T;Norose K;Yano A.;永井宏樹
- 通讯作者:永井宏樹
Screening of novel protein substrates of the Legionella Dot/Icm type IV secretion system based on the properties of C-terminal translocation signals
基于C端易位信号特性筛选军团菌Dot/Icm IV型分泌系统的新型蛋白底物
- DOI:
- 发表时间:2006
- 期刊:
- 影响因子:0
- 作者:川本;進;Hiroki Nagai
- 通讯作者:Hiroki Nagai
Molecular analysis of Legionella type IV effectors..
IV 型军团菌效应子的分子分析..
- DOI:
- 发表时间:2008
- 期刊:
- 影响因子:0
- 作者:喜多 英二;東 伸岳;岡山 明子;百武晃宏;久堀智子;Akihiro Hyakutake;Tomoko Kubori;Hiroki Nagai
- 通讯作者:Hiroki Nagai
IV型分泌装置の機能または自己分泌能を失った変異型DotAの単離と解析
失去IV型分泌器功能或自分泌能力的突变体DotA的分离和分析
- DOI:
- 发表时间:2008
- 期刊:
- 影响因子:0
- 作者:喜多 英二;東 伸岳;岡山 明子;百武晃宏
- 通讯作者:百武晃宏
Isolation and characterization of DotA mutants defective either in DotA secretion or in DotA function
DotA 分泌或 DotA 功能缺陷的 DotA 突变体的分离和表征
- DOI:
- 发表时间:2008
- 期刊:
- 影响因子:0
- 作者:喜多 英二;東 伸岳;岡山 明子;百武晃宏;久堀智子;Akihiro Hyakutake
- 通讯作者:Akihiro Hyakutake
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NAGAI Hiroki其他文献
液固比バッチ試験によるヒ素の吸脱着反応の評価
液固比批量试验评价砷吸附/解吸反应
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2021 - 期刊:
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FUJITA Koji;KURANE Kakeru;TAKAHASHI Koichi;NAGAI Hiroki;奈佐原寅太郎 - 通讯作者:
奈佐原寅太郎
Experiments and Dynamical Threshold of Bubble Nucleus Growth in Water by Non-Recirculating Cavitation Tunnel
非循环空化隧道水中气泡核生长的实验及动态阈值
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10.11395/jjsem.22.246 - 发表时间:
2023 - 期刊:
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- 作者:
IKAMI Tsubasa;FUJITA Koji;NAGAI Hiroki;MATSUDA Yu;EGAMI Yasuhiro;藤川俊秀,江頭竜,陣内楓,藤川重雄 - 通讯作者:
藤川俊秀,江頭竜,陣内楓,藤川重雄
Effects of Propeller Position and Rotation Direction on the Ishii Wing at a Low Reynolds Number
低雷诺数下螺旋桨位置和旋转方向对石井翼的影响
- DOI:
10.2322/tjsass.64.22 - 发表时间:
2021 - 期刊:
- 影响因子:1.1
- 作者:
FUJITA Koji;KURANE Kakeru;TAKAHASHI Koichi;NAGAI Hiroki - 通讯作者:
NAGAI Hiroki
Thermal state estimation based on Assisted Ensemble Kalman Filter
基于辅助系综卡尔曼滤波器的热状态估计
- DOI:
10.1299/transjsme.21-00010 - 发表时间:
2021 - 期刊:
- 影响因子:0
- 作者:
TANAKA Hiroto;MISAKA Takashi;FUJITA Koji;NAGAI Hiroki - 通讯作者:
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Investigation of Data Acquisition Conditions for Dynamic Mode Decomposition in Unsteady PSP Measurement
非稳态 PSP 测量中动态模式分解的数据采集条件研究
- DOI:
10.3154/tvsj.41.11 - 发表时间:
2021 - 期刊:
- 影响因子:0
- 作者:
IKAMI Tsubasa;FUJITA Koji;NAGAI Hiroki;MATSUDA Yu;EGAMI Yasuhiro - 通讯作者:
EGAMI Yasuhiro
NAGAI Hiroki的其他文献
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{{ truncateString('NAGAI Hiroki', 18)}}的其他基金
Molecular basis of evasion of Legionella from xenophagy
军团菌逃避异体吞噬的分子基础
- 批准号:
26670212 - 财政年份:2014
- 资助金额:
$ 2.57万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Key technology Research and High-altitude demonstration to realize the world first Mars exploration using Airplane
实现世界首次飞机火星探测关键技术研究及高空论证
- 批准号:
24246136 - 财政年份:2012
- 资助金额:
$ 2.57万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
T4SS supermolecular complex containing core complex and ATPase
含有核心复合物和ATP酶的T4SS超分子复合物
- 批准号:
24659198 - 财政年份:2012
- 资助金额:
$ 2.57万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Molecular disection of type IVB secretion system core complex.
IVB 型分泌系统核心复合体的分子解剖。
- 批准号:
23390105 - 财政年份:2011
- 资助金额:
$ 2.57万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
High-accuracy estimation of aerodynamic heating in high enthalpy flowusing Temperature-Sensitive Paint and Inverse method
利用温敏涂料和反演法高精度估算高焓流中的气动加热
- 批准号:
20760545 - 财政年份:2008
- 资助金额:
$ 2.57万 - 项目类别:
Grant-in-Aid for Young Scientists (B)
Molecular analyses of a Legionella ubiquitin ligase.
军团菌泛素连接酶的分子分析。
- 批准号:
19590446 - 财政年份:2007
- 资助金额:
$ 2.57万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
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