The T4bSS of Legionella features a two-step secretion pathway with an inner membrane intermediate for secretion of transmembrane effectors

被引:0
作者
Malmsheimer, Silke [1 ]
Grin, Iwan [1 ]
Bohn, Erwin [2 ]
Franz-Wachtel, Mirita [3 ]
Macek, Boris [3 ]
Sahr, Tobias [4 ]
Smollich, Fabian [1 ]
Chetrit, David [5 ]
Meir, Amit [5 ,6 ,7 ]
Roy, Craig [5 ]
Buchrieser, Carmen [4 ]
Wagner, Samuel [1 ,8 ,9 ]
机构
[1] Univ Tubingen, Interfac Inst Microbiol & Infect Med IMIT, Sect Cellular & Mol Microbiol, Tubingen, Germany
[2] Univ Tubingen, Inst Med Microbiol & Hyg, Interfac Inst Microbiol & Infect Med IMIT, Tubingen, Germany
[3] Univ Tubingen, Proteome Ctr Tubingen, Tubingen, Germany
[4] Univ Paris Cite, Inst Pasteur, Biol Bacteries Intracellulaires, Paris, France
[5] Yale Univ, Dept Microbial Pathogenesis, New Haven, CT USA
[6] Birkbeck Inst Struct & Mol Biol, London, England
[7] UCL, London, England
[8] Excellence ClusterControlling Microbes Fight Infec, Tubingen, Germany
[9] German Ctr Infect Res DZIF, Partner Site Tubingen, Tubingen, Germany
基金
欧洲研究理事会;
关键词
PROTEIN; PNEUMOPHILA; IDENTIFICATION; SYSTEM; TOPOLOGY; COMPLEX; DOTA;
D O I
10.1371/journal.ppat.1012118
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
To promote intracellular survival and infection, Legionella spp. translocate hundreds of effector proteins into eukaryotic host cells using a type IV b protein secretion system (T4bSS). T4bSS are well known to translocate soluble as well as transmembrane domain-containing effector proteins (TMD-effectors) but the mechanisms of secretion are still poorly understood. Herein we investigated the secretion of hydrophobic TMD-effectors, of which about 80 were previously reported to be encoded by L. pneumophila. A proteomic analysis of fractionated membranes revealed that TMD-effectors are targeted to and inserted into the bacterial inner membranes of L. pneumophila independent of the presence of a functional T4bSS. While the T4bSS chaperones IcmS and IcmW were critical for secretion of all tested TMD-effectors, they did not influence inner membrane targeting of these proteins. As for soluble effector proteins, translocation of all investigated TMD-effectors depended on a C-terminal secretion signal. A deeper analysis of the TMD-effector SidF showed that this signal needed to be presented towards the cytoplasmic side of the inner membrane and that a small periplasmic loop was required for efficient translocation. We propose that strongly hydrophobic TMD-effectors are secreted in a two-step secretion process: Initially, an inner membrane intermediate is formed, that is extracted towards the cytoplasmic side, possibly by the help of the type IV coupling protein complex and subsequently secreted into eukaryotic host cells by the T4bSS core complex. Overall, our study highlights the amazing versatility of T4bSS to secrete soluble and TMD-effectors from different subcellular locations of the bacterial cell.
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页数:21
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