Comprehensive identification of protein substrates of the Dot/Icm type IV transporter of Legionella pneumophila.
Comprehensive identification of protein substrates of the Dot/Icm type IV transporter of Legionella pneumophila.
复制标题
DOI：
10.1371/journal.pone.0017638
复制
发表时间：
2011-03-09
期刊：
影响因子：
3.7
通讯作者：
Luo ZQ
中科院分区：
文献类型：
--
作者：
Zhu W;Banga S;Tan Y;Zheng C;Stephenson R;Gately J;Luo ZQ
嗜肺军团菌Dot/Icm系统转移的大量蛋白质已通过各种策略鉴定。毫无例外，这些策略基于与测试蛋白质相关的一个或多个特征。鉴于所鉴定的蛋白质表现出高度的多样性，这些筛选中可能遗漏了一些底物。本研究采用系统调查的方法，对我国北方地区的南方红豆杉进行了调查。pneumophila基因组通过测试大于300个碱基对的假设ORFs的Dot/Icm依赖性易位。832个ORFs中有798个成功融合到β-内酰胺酶的羧基端。使用β-内酰胺酶报告底物CCF 4-AM确定融合物转移到哺乳动物细胞中。这些努力导致了164个蛋白质易位阳性的鉴定。其中，70种蛋白质是Dot/Icm系统的新底物。这些结果使实验确认的Dot/Icm基板的总数达到275。这些鉴定的蛋白质的C-末端的序列分析显示，Lpg 2844，其中包含已知是重要的点/Icm依赖的蛋白质转移的几个功能，可以以高效率易位。因此，我们的努力已经确定了大量的新底物的点/Icm系统，并揭示了可识别的这种蛋白质转运蛋白的不同功能。
A large number of proteins transferred by the Legionella pneumophila Dot/Icm system have been identified by various strategies. With no exceptions, these strategies are based on one or more characteristics associated with the tested proteins. Given the high level of diversity exhibited by the identified proteins, it is possible that some substrates have been missed in these screenings. In this study, we took a systematic method to survey the L. pneumophila genome by testing hypothetical orfs larger than 300 base pairs for Dot/Icm-dependent translocation. 798 of the 832 analyzed orfs were successfully fused to the carboxyl end of β-lactamase. The transfer of the fusions into mammalian cells was determined using the β-lactamase reporter substrate CCF4-AM. These efforts led to the identification of 164 proteins positive in translocation. Among these, 70 proteins are novel substrates of the Dot/Icm system. These results brought the total number of experimentally confirmed Dot/Icm substrates to 275. Sequence analysis of the C-termini of these identified proteins revealed that Lpg2844, which contains few features known to be important for Dot/Icm-dependent protein transfer can be translocated at a high efficiency. Thus, our efforts have identified a large number of novel substrates of the Dot/Icm system and have revealed the diverse features recognizable by this protein transporter.
登录
查看更多内容
DOI：
10.1084/jem.158.6.2108
发表时间：
1983-12-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Horwitz MA
通讯作者：
Horwitz MA
DOI：
10.1073/pnas.0907200106
发表时间：
2009-08-18
影响因子：
11.1
作者：
Ge, Jianning;Xu, Hao;Shao, Feng
通讯作者：
Shao, Feng
影响因子：
3.6
作者：
Bardill, JP;Miller, JL;Vogel, JP
通讯作者：
Vogel, JP
影响因子：
3.6
作者：
Kubori, Tomoko;Hyakutake, Akihiro;Nagai, Hiroki
通讯作者：
Nagai, Hiroki
影响因子：
3.4
作者：
Lomma, M.;Dervins-Ravault, D.;Buchrieser, C.
通讯作者：
Buchrieser, C.