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The proteins DotY and DotZ modulate the dynamics and localization of the type IVB coupling complex of Legionella pneumophila

DOI: 10.1111/mmi.14847 DOI Help

Authors: Kevin Mace (Institute of Structural and Molecular Biology, Birkbeck and UCL) , Amit Meir (Institute of Structural and Molecular Biology, Birkbeck and UCL; Yale University) , Natalya Lukoyanova (Institute of Structural and Molecular Biology, Birkbeck and UCL) , Luying Liu (Yale University) , David Chetrit (Yale University) , Manuela K. Hospenthal (Institute of Structural and Molecular Biology, Birkbeck and UCL; ETH Z├╝rich) , Craig R. Roy (Yale University) , Gabriel Waksman (Institute of Structural and Molecular Biology, Birkbeck and UCL)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Molecular Microbiology

State: Published (Approved)
Published: November 2021
Diamond Proposal Number(s): 14704

Abstract: Legionella pneumophila is an opportunistic pathogen infecting alveolar macrophages and protozoa species. Legionella utilizes a Type IV Secretion System (T4SS) to translocate over 300 effector proteins into its host cell. In a recent study, we isolated and solved the cryo-EM structure of the Type IV Coupling Complex (T4CC), a large cytoplasmic determinant associated with the inner membrane that recruits effector proteins for delivery to the T4SS for translocation. The T4CC is composed of a DotLMNYZ hetero-pentameric core from which the flexible IcmSW module flexibly protrudes. The DotY and DotZ proteins were newly reported members of this complex and their role remained elusive. In this study, we observed the effect of deleting DotY and DotZ on T4CC stability and localization. Furthermore, we found these two proteins are co-dependent, whereby the deletion of DotY resulted in DotZ absence from the coupling complex, and vice versa. Additional cryo-EM data analysis revealed the dynamic movement of the IcmSW module is modified by the DotY/Z proteins. We therefore determined the likely function of DotY and DotZ and revealed their importance on T4CC function.

Diamond Keywords: Bacteria; Legionnaires' Disease

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: Krios II-Titan Krios II at Diamond

Added On: 26/11/2021 10:26

Discipline Tags:

Pathogens Infectious Diseases Health & Wellbeing Structural biology Life Sciences & Biotech

Technical Tags:

Microscopy Electron Microscopy (EM) Cryo Electron Microscopy (Cryo EM)