Atomic Structure of T6SS Reveals Interlaced Array Essential to Function

被引：127

作者：

Clemens, Daniel L. ^{[1
]}

Ge, Peng ^{[2
,3
]}

Lee, Bai-Yu ^{[1
]}

Horwitz, Marcus A. ^{[1
,2
]}

Zhou, Z. Hong ^{[2
,3
]}

机构：

[1] Univ Calif Los Angeles, Dept Med, Los Angeles, CA 90095 USA

[2] Univ Calif Los Angeles, Dept Microbiol Immunol & Mol Genet, Los Angeles, CA 90095 USA

[3] Univ Calif Los Angeles, Calif NanoSyst Inst, Los Angeles, CA 90095 USA

来源：

CELL | 2015年 / 160卷 / 05期

基金：

美国国家卫生研究院; 美国国家科学基金会;

关键词：

VI SECRETION SYSTEM; FRANCISELLA-TULARENSIS; PSEUDOMONAS-AERUGINOSA; RESOLUTION; PHAGE; BACTERIOPHAGE-T4; IDENTIFICATION; MACROPHAGES; TRAFFICKING; MECHANISM;

D O I：

10.1016/j.cell.2015.02.005

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Type VI secretion systems (T6SSs) are newly identified contractile nanomachines that translocate effector proteins across bacterial membranes. The Francisella pathogenicity island, required for bacterial phagosome escape, intracellular replication, and virulence, was presumed to encode a T6SS-like apparatus. Here, we experimentally confirm the identity of this T6SS and, by cryo electron microscopy (cryoEM), show the structure of its post-contraction sheath at 3.7 angstrom resolution. We demonstrate the assembly of this T6SS by IglA/IglB and secretion of its putative effector proteins in response to environmental stimuli. The sheath has a quaternary structure with handedness opposite that of contracted sheath of T4 phage tail and is organized in an interlaced two-dimensional array by means of beta sheet augmentation. By structure-based mutagenesis, we show that this interlacing is essential to secretion, phagosomal escape, and intracellular replication. Our atomic model of the T6SS will facilitate design of drugs targeting this highly prevalent secretion apparatus.

引用

页码：940 / 951

页数：12

共 49 条

[1] PHENIX: a comprehensive Python']Python-based system for macromolecular structure solution [J].

Adams, Paul D. ;

Afonine, Pavel V. ;

Bunkoczi, Gabor ;

Chen, Vincent B. ;

Davis, Ian W. ;

Echols, Nathaniel ;

Headd, Jeffrey J. ;

Hung, Li-Wei ;

Kapral, Gary J. ;

Grosse-Kunstleve, Ralf W. ;

McCoy, Airlie J. ;

Moriarty, Nigel W. ;

Oeffner, Robert ;

Read, Randy J. ;

Richardson, David C. ;

Richardson, Jane S. ;

Terwilliger, Thomas C. ;

Zwart, Peter H. .

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2010, 66 :213-221

[2] Structural Conservation of the Myoviridae Phage Tail Sheath Protein Fold [J].

Aksyuk, Anastasia A. ;

Kurochkina, Lidia P. ;

Fokine, Andrei ;

Forouhar, Farhad ;

Mesyanzhinov, Vadim V. ;

Tong, Liang ;

Rossmann, Michael G. .

STRUCTURE, 2011, 19 (12) :1885-1894

[3] The tail sheath structure of bacteriophage T4: a molecular machine for infecting bacteria [J].

Aksyuk, Anastasia A. ;

Leiman, Petr G. ;

Kurochkina, Lidia P. ;

Shneider, Mikhail M. ;

Kostyuchenko, Victor A. ;

Mesyanzhinov, Vadim V. ;

Rossmann, Michael G. .

EMBO JOURNAL, 2009, 28 (07) :821-829

[4] The Francisella tularensis pathogenicity island encodes a secretion system that is required for phagosome escape and virulence [J].

Barker, Jeffrey R. ;

Chong, Audrey ;

Wehrly, Tara D. ;

Yu, Jieh-Juen ;

Rodriguez, Stephen A. ;

Liu, Jirong ;

Celli, Jean ;

Arulanandam, Bernard P. ;

Klose, Karl E. .

MOLECULAR MICROBIOLOGY, 2009, 74 (06) :1459-1470

[5] Type VI secretion requires a dynamic contractile phage tail-like structure [J].

Basler, M. ;

Pilhofer, M. ;

Henderson, G. P. ;

Jensen, G. J. ;

Mekalanos, J. J. .

NATURE, 2012, 483 (7388) :182-U78

[6] Tit-for-Tat: Type VI Secretion System Counterattack during Bacterial Cell-Cell Interactions [J].

Basler, Marek ;

Ho, Brian T. ;

Mekalanos, John J. .

CELL, 2013, 152 (04) :884-894

[7] Version 1.2 of the Crystallography and NMR system [J].

Brunger, Axel T. .

NATURE PROTOCOLS, 2007, 2 (11) :2728-2733

[8] In vivo and in vitro protein solubility assays using split GFP [J].

Cabantous, Stephanie ;

Waldo, Geoffrey S. .

NATURE METHODS, 2006, 3 (10) :845-854

[9] Movies of Ice-Embedded Particles Enhance Resolution in Electron Cryo-Microscopy [J].

Campbell, Melody G. ;

Cheng, Anchi ;

Brilot, Axel F. ;

Moeller, Arne ;

Lyumkis, Dmitry ;

Veesler, David ;

Pan, Junhua ;

Harrison, Stephen C. ;

Potter, Clinton S. ;

Carragher, Bridget ;

Grigorieff, Nikolaus .

STRUCTURE, 2012, 20 (11) :1823-1828

[10] Autophagy-mediated reentry of Francisella tularensis into the endocytic compartment after cytoplasmic replication [J].

Checroun, Claire ;

Wehrly, Tara D. ;

Fischer, Elizabeth R. ;

Hayes, Stanley F. ;

Celli, Jean .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (39) :14578-14583

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