Legionnaires' disease is caused by a lethal colonization of alveolar macrophages with the Gram-negative bacterium Legionella pneumophila. LpGT (L. pneumophila glucosyltransferase; also known as Lgt1) has recently been identified as a virulence factor, shutting down protein synthesis in the human cell by specific glucosylation of EF1A (elongation factor 1A), using an unknown mode of substrate recognition and a retaining mechanism for glycosyl transfer. We have determined the crystal structure of LpGT in complex with substrates, revealing a GT-A fold with two unusual protruding domains. Through structure-guided mutagenesis of LpGT, several residues essential for binding of the UDP-glucose-donor and EF1A-acceptor substrates were identified, which also affected L. pneumophila virulence as demonstrated by microinjection studies. Together, these results suggested that a positively charged EF1A loop binds to a negatively charged conserved groove on the LpGT structure, and that two asparagine residues are essential for catalysis. Furthermore, we showed that two further L. pneumophila glycosyltransferases possessed the conserved UDP-glucose-binding sites and EF1A-binding grooves, and are, like LpGT, translocated into the macrophage through the Icm/Dot (intracellular multiplication/defect in organelle trafficking) system.
Skip Nav Destination
Article navigation
March 2010
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
February 24 2010
Molecular mechanism of elongation factor 1A inhibition by a Legionella pneumophila glycosyltransferase
Ramon Hurtado-Guerrero;
Ramon Hurtado-Guerrero
1
*Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, U.K.
1Correspondence may be addressed to either of these authors (email R.HurtadoGuerrero@dundee.ac.uk or dmfvanaalten@dundee.ac.uk).
Search for other works by this author on:
Tal Zusman;
Tal Zusman
†Molecular Microbiology and Biotechnology, Life Sciences, Tel Aviv University, Tel Aviv, Israel
Search for other works by this author on:
Shalini Pathak;
Shalini Pathak
*Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, U.K.
Search for other works by this author on:
Adel F. M. Ibrahim;
Adel F. M. Ibrahim
‡DNA Manipulation Team, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, U.K.
Search for other works by this author on:
Sharon Shepherd;
Sharon Shepherd
*Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, U.K.
Search for other works by this author on:
Alan Prescott;
Alan Prescott
§Division of Cell Biology and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, U.K.
Search for other works by this author on:
Gil Segal;
Gil Segal
†Molecular Microbiology and Biotechnology, Life Sciences, Tel Aviv University, Tel Aviv, Israel
Search for other works by this author on:
Daan M. F. van Aalten
Daan M. F. van Aalten
1
*Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, U.K.
1Correspondence may be addressed to either of these authors (email R.HurtadoGuerrero@dundee.ac.uk or dmfvanaalten@dundee.ac.uk).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
September 01 2009
Revision Received:
December 22 2009
Accepted:
December 23 2009
Accepted Manuscript online:
December 23 2009
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2010 Biochemical Society
2010
Biochem J (2010) 426 (3): 281–292.
Article history
Received:
September 01 2009
Revision Received:
December 22 2009
Accepted:
December 23 2009
Accepted Manuscript online:
December 23 2009
Citation
Ramon Hurtado-Guerrero, Tal Zusman, Shalini Pathak, Adel F. M. Ibrahim, Sharon Shepherd, Alan Prescott, Gil Segal, Daan M. F. van Aalten; Molecular mechanism of elongation factor 1A inhibition by a Legionella pneumophila glycosyltransferase. Biochem J 15 March 2010; 426 (3): 281–292. doi: https://doi.org/10.1042/BJ20091351
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.