한빛사 논문
Debnath Ghosala, Kwangcheol C. Jeongb,c, Yi-Wei Changa, Jacob Gyoreb, Lin Tengc, Adam Gardnerd, Joseph P. Vogelb,* & Grant J. Jensena,e,*
aDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA
bDepartment of Molecular Microbiology, Washington University School of Medicine, St Louis, MO, USA
cDepartment of Animal Sciences & Emerging Pathogens Institute, University of Florida, Gainesville, FL, USA
dMolecular Graphics Laboratory, Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA, USA
eHoward Hughes Medical Institute, California Institute of Technology, Pasadena, CA, USA
These authors contributed equally: Debnath Ghosal, Kwangcheol C. Jeong.
*Correspondence to Joseph P. Vogel or Grant J. Jensen.
Abstract
Legionella pneumophila survives and replicates inside host cells by secreting ~300 effectors through the defective in organelle trafficking (Dot)/intracellular multiplication (Icm) type IVB secretion system (T4BSS). Here, we used complementary electron cryotomography and immunofluorescence microscopy to investigate the molecular architecture and biogenesis of the Dot/Icm secretion apparatus. Electron cryotomography mapped the location of the core and accessory components of the Legionella core transmembrane subcomplex, revealing a well-ordered central channel that opens into a large, windowed secretion chamber with an unusual 13-fold symmetry. Immunofluorescence microscopy deciphered an early-stage assembly process that begins with the targeting of Dot/Icm components to the bacterial poles. Polar targeting of this T4BSS is mediated by two Dot/Icm proteins, DotU and IcmF, that, interestingly, are homologues of the T6SS membrane complex components TssL and TssM, suggesting that the Dot/Icm T4BSS is a hybrid system. Together, these results revealed that the Dot/Icm complex assembles in an ‘axial-to-peripheral’ pattern.
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