Activation of Ran GTPase by a Legionella effector promotes microtubule polymerization, pathogen vacuole motility and infection
Standard
Activation of Ran GTPase by a Legionella effector promotes microtubule polymerization, pathogen vacuole motility and infection. / Rothmeier, Eva; Pfaffinger, Gudrun; Hoffmann, Christine; Harrison, Christopher F; Grabmayr, Heinrich; Repnik, Urska; Hannemann, Mandy; Wölke, Stefan; Bausch, Andreas; Griffiths, Gareth; Müller-Taubenberger, Annette; Itzen, Aymelt; Hilbi, Hubert.
In: PLOS PATHOG, Vol. 9, No. 9, 09.2013, p. e1003598.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - Activation of Ran GTPase by a Legionella effector promotes microtubule polymerization, pathogen vacuole motility and infection
AU - Rothmeier, Eva
AU - Pfaffinger, Gudrun
AU - Hoffmann, Christine
AU - Harrison, Christopher F
AU - Grabmayr, Heinrich
AU - Repnik, Urska
AU - Hannemann, Mandy
AU - Wölke, Stefan
AU - Bausch, Andreas
AU - Griffiths, Gareth
AU - Müller-Taubenberger, Annette
AU - Itzen, Aymelt
AU - Hilbi, Hubert
PY - 2013/9
Y1 - 2013/9
N2 - The causative agent of Legionnaires' disease, Legionella pneumophila, uses the Icm/Dot type IV secretion system (T4SS) to form in phagocytes a distinct "Legionella-containing vacuole" (LCV), which intercepts endosomal and secretory vesicle trafficking. Proteomics revealed the presence of the small GTPase Ran and its effector RanBP1 on purified LCVs. Here we validate that Ran and RanBP1 localize to LCVs and promote intracellular growth of L. pneumophila. Moreover, the L. pneumophila protein LegG1, which contains putative RCC1 Ran guanine nucleotide exchange factor (GEF) domains, accumulates on LCVs in an Icm/Dot-dependent manner. L. pneumophila wild-type bacteria, but not strains lacking LegG1 or a functional Icm/Dot T4SS, activate Ran on LCVs, while purified LegG1 produces active Ran(GTP) in cell lysates. L. pneumophila lacking legG1 is compromised for intracellular growth in macrophages and amoebae, yet is as cytotoxic as the wild-type strain. A downstream effect of LegG1 is to stabilize microtubules, as revealed by conventional and stimulated emission depletion (STED) fluorescence microscopy, subcellular fractionation and Western blot, or by microbial microinjection through the T3SS of a Yersinia strain lacking endogenous effectors. Real-time fluorescence imaging indicates that LCVs harboring wild-type L. pneumophila rapidly move along microtubules, while LCVs harboring ΔlegG1 mutant bacteria are stalled. Together, our results demonstrate that Ran activation and RanBP1 promote LCV formation, and the Icm/Dot substrate LegG1 functions as a bacterial Ran activator, which localizes to LCVs and promotes microtubule stabilization, LCV motility as well as intracellular replication of L. pneumophila.
AB - The causative agent of Legionnaires' disease, Legionella pneumophila, uses the Icm/Dot type IV secretion system (T4SS) to form in phagocytes a distinct "Legionella-containing vacuole" (LCV), which intercepts endosomal and secretory vesicle trafficking. Proteomics revealed the presence of the small GTPase Ran and its effector RanBP1 on purified LCVs. Here we validate that Ran and RanBP1 localize to LCVs and promote intracellular growth of L. pneumophila. Moreover, the L. pneumophila protein LegG1, which contains putative RCC1 Ran guanine nucleotide exchange factor (GEF) domains, accumulates on LCVs in an Icm/Dot-dependent manner. L. pneumophila wild-type bacteria, but not strains lacking LegG1 or a functional Icm/Dot T4SS, activate Ran on LCVs, while purified LegG1 produces active Ran(GTP) in cell lysates. L. pneumophila lacking legG1 is compromised for intracellular growth in macrophages and amoebae, yet is as cytotoxic as the wild-type strain. A downstream effect of LegG1 is to stabilize microtubules, as revealed by conventional and stimulated emission depletion (STED) fluorescence microscopy, subcellular fractionation and Western blot, or by microbial microinjection through the T3SS of a Yersinia strain lacking endogenous effectors. Real-time fluorescence imaging indicates that LCVs harboring wild-type L. pneumophila rapidly move along microtubules, while LCVs harboring ΔlegG1 mutant bacteria are stalled. Together, our results demonstrate that Ran activation and RanBP1 promote LCV formation, and the Icm/Dot substrate LegG1 functions as a bacterial Ran activator, which localizes to LCVs and promotes microtubule stabilization, LCV motility as well as intracellular replication of L. pneumophila.
KW - Animals
KW - Bacterial Proteins
KW - Cell Line
KW - Enzyme Activation
KW - GTPase-Activating Proteins
KW - Gene Silencing
KW - Humans
KW - Legionella pneumophila
KW - Legionnaires' Disease
KW - Macrophages
KW - Mice
KW - Microtubule Proteins
KW - Microtubules
KW - Mutation
KW - Phagocytosis
KW - Phagosomes
KW - Polymerization
KW - Protein Stability
KW - Protein Transport
KW - Virus Replication
KW - ran GTP-Binding Protein
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1371/journal.ppat.1003598
DO - 10.1371/journal.ppat.1003598
M3 - SCORING: Journal article
C2 - 24068924
VL - 9
SP - e1003598
JO - PLOS PATHOG
JF - PLOS PATHOG
SN - 1553-7366
IS - 9
ER -