Protein LidA from Legionella is a Rab GTPase supereffector

Schoebel, Stefan; Cichy, Adam Leszek; Goody, Roger S.; Itzen, Aymelt

doi:10.1073/pnas.1113133108

Cited by 75 publications

(91 citation statements)

References 38 publications

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“…Motivated by recent findings, we investigated the disruption of GDI:Rab:GDP complexes by proteins with high affinity to GDP-bound Rabs. The Legionella protein LidA has a K d for Rab1 in the subnanomolar range (15) and based on a comparison of the binding surfaces of LidA and GDI on Rabs should therefore efficiently compete with GDI-binding to Rab1b, which is of nanomolar affinity (3). We confirmed this hypothesis by displacing GDI from the complex with Rab1b-f-NBD (50 nM) by addition of LidA (1 μM) (Fig.…”

Section: Resultsmentioning

confidence: 99%

Posttranslational modifications of Rab proteins cause effective displacement of GDP dissociation inhibitor

Oesterlin

Goody

Itzen

2012

Proc. Natl. Acad. Sci. U.S.A.

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Intracellular vesicular trafficking is regulated by approximately 60 members of the Rab subfamily of small Ras-like GDP/GTP binding proteins. Rab proteins cycle between inactive and active states as well as between cytosolic and membrane bound forms. Membrane extraction/delivery and cytosolic distribution of Rabs is mediated by interaction with the protein GDP dissociation inhibitor (GDI) that binds to prenylated inactive (GDP-bound) Rab proteins. Because the Rab:GDP:GDI complex is of high affinity, the question arises of how GDI can be displaced efficiently from Rab protein in order to allow the necessary recruitment of the Rab to its specific target membrane. While there is strong evidence that DrrA, as a bacterially encoded GDP/GTP exchange factor, contributes to this event, we show here that posttranslational modifications of Rabs can also modulate the affinity for GDI and thus cause effective displacement of GDI from Rab:GDI complexes. These activities have been found associated with the phosphocholination and adenylylation activities of the enzymes AnkX and DrrA/SidM, respectively, from the pathogenic bacterium Legionella pneumophila. Both modifications occur after spontaneous dissociation of Rab:GDI complexes within their natural equilibrium. Therefore, the effective GDI displacement that is observed is caused by inhibition of reformation of Rab:GDI complexes. Interestingly, in contrast to adenylylation by DrrA, AnkX can covalently modify inactive Rabs with high catalytic efficiency even when GDP is bound to the GTPase and hence can inhibit binding of GDI to Rab:GDP complexes. We therefore speculate that human cells could employ similar mechanisms in the absence of infection to effectively displace Rabs from GDI.T he intracellular activity of most signaling proteins is regulated at the temporal and spatial level. In the regulation of vesicular trafficking, small Ras-like GTPases of the Rab subfamily are controlled in their distribution between the cytosol and intracellular membranes, which is in turn coupled to the binding to either GDP or GTP. Cycling between the cytosol and membranes is dependent on the activation state of the Rab protein. In the active GTP-bound form, Rabs are associated with membranes, where they interact with effector proteins that promote diverse steps in vesicular trafficking. When bound to GDP, Rabs are defined as inactive and are predominantly distributed in the cytosol bound to GDI. In order to associate with the cytosolic surface of intracellular membranes, most Rab proteins possess two hydrophobic prenyl (geranylgeranyl) moieties at their structurally flexible C terminus that are posttranslationally attached via the concerted action of Rab geranylgeranyl transferase (RabGGTase) and Rab escort protein utilizing geranylgeranyl pyrophosphate as a cosubstrate. These geranylgeranyl groups mediate stable peripheral binding of Rabs to a cognate membrane by inserting the hydrophobic anchors into the lipid bilayer (1). Activation of Rab proteins is catalyzed by GDP/GTP exchange factors...

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Section: Resultsmentioning

confidence: 99%

Posttranslational modifications of Rab proteins cause effective displacement of GDP dissociation inhibitor

Oesterlin

Goody

Itzen

2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Furthermore, the covalent modification has been shown to inhibit binding of GDI, thereby not allowing extraction from the LCV membrane (24). During this entrapment phase, the supereffector LidA might act as a tethering factor for endoplasmic reticulum-derived vesicles because it is still capable of binding AMP-Rab1 with high affinity and is present at the LCV within 20 min after infection (20,53). In contrast to LidA from Legionella, binding of the human effector protein Mical is not possible in the modified state.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Enzymes from Legionella pneumophila Involved in Reversible Adenylylation of Rab1 Protein

Müller

Shkumatov²,

Oesterlin

et al. 2012

Journal of Biological Chemistry

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Background: Covalent modification of small GTPases by pathogens is an emerging field in current research. Results: Quantitative analysis of effects, kinetics, and substrate specificities of adenylylation by DrrA and deadenylylation by SidD was performed. Conclusion: Adenylylation and deadenylylation are means to tightly regulate Rab1 function by Legionella proteins. Significance: This study increases our understanding of Legionella pneumophila subverting Rab protein function during infection.

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“…Interestingly, there is a secreted Legionella protein that has some of the required properties of a GTPase effector and localizes to the LCV surface (Conover et al, 2003). However, the affinity between Rab1 and LidA is so high that it could not be measured, neither in the GTP nor the GDP form (Schoebel et al, 2011). In particular, the dissociation rate constants of the complexes are extremely slow.…”

Section: Learning About Vesicular Transport From Bacteriamentioning

confidence: 99%

“…In particular, the dissociation rate constants of the complexes are extremely slow. Rab8 is also bound extremely tightly, and the structure of the LidA : Rab8 : GppNHp complex was determined, revealing a very large interaction surface area (Schoebel et al, 2011), as also seen in the corresponding Rab1 complex (Cheng et al, 2012). Rab6 is bound more weakly, thus allowing the affinity to be measured.…”

Section: Learning About Vesicular Transport From Bacteriamentioning

confidence: 99%

Mechanisms of action of Rab proteins, key regulators of intracellular vesicular transport

Goody

Müller

2016

Biological Chemistry

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Our understanding of the manner in which Rab proteins regulate intracellular vesicular transport has progressed remarkably in the last one or two decades by application of a wide spectrum of biochemical, biophysical and cell biological methods, augmented by the methods of chemical biology. Important additional insights have arisen from examination of the manner in which certain bacteria can manipulate vesicular transport mechanisms. The progress in these areas is summarized here.

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Protein LidA from Legionella is a Rab GTPase supereffector

Cited by 75 publications

References 38 publications

Posttranslational modifications of Rab proteins cause effective displacement of GDP dissociation inhibitor

Posttranslational modifications of Rab proteins cause effective displacement of GDP dissociation inhibitor

Characterization of Enzymes from Legionella pneumophila Involved in Reversible Adenylylation of Rab1 Protein

Mechanisms of action of Rab proteins, key regulators of intracellular vesicular transport

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