The assembly of a GTPase-kinase signalling complex by a bacterial catalytic scaffold

Andrey S. Selyunin, Sarah E. Sutton, Bethany A. Weigele, L. Evan Reddick, Robert C. Orchard, Stefan M. Bresson, Diana R. Tomchick, Neal M. Alto

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

The fidelity and specificity of information flow within a cell is controlled by scaffolding proteins that assemble and link enzymes into signalling circuits. These circuits can be inhibited by bacterial effector proteins that post-translationally modify individual pathway components. However, there is emerging evidence that pathogens directly organize higher-order signalling networks through enzyme scaffolding, and the identity of the effectors and their mechanisms of action are poorly understood. Here we identify the enterohaemorrhagic Escherichia coli O157:H7 type III effector EspG as a regulator of endomembrane trafficking using a functional screen, and report ADP-ribosylation factor (ARF) GTPases and p21-activated kinases (PAKs) as its relevant host substrates. The 2.5 crystal structure of EspG in complex with ARF6 shows how EspG blocks GTPase-activating-protein-assisted GTP hydrolysis, revealing a potent mechanism of GTPase signalling inhibition at organelle membranes. In addition, the 2.8 crystal structure of EspG in complex with the autoinhibitory Iα3-helix of PAK2 defines a previously unknown catalytic site in EspG and provides an allosteric mechanism of kinase activation by a bacterial effector. Unexpectedly, ARF and PAKs are organized on adjacent surfaces of EspG, indicating its role as a catalytic scaffold that effectively reprograms cellular events through the functional assembly of GTPase-kinase signalling complex.

Original languageEnglish (US)
Pages (from-to)107-113
Number of pages7
JournalNature
Volume469
Issue number7328
DOIs
StatePublished - Jan 6 2011

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GTP Phosphohydrolases
ADP-Ribosylation Factors
p21-Activated Kinases
Phosphotransferases
Enterohemorrhagic Escherichia coli
GTPase-Activating Proteins
Escherichia coli O157
Bacterial Proteins
Enzymes
Guanosine Triphosphate
Organelles
Catalytic Domain
Hydrolysis
Membranes

ASJC Scopus subject areas

  • General

Cite this

Selyunin, A. S., Sutton, S. E., Weigele, B. A., Reddick, L. E., Orchard, R. C., Bresson, S. M., ... Alto, N. M. (2011). The assembly of a GTPase-kinase signalling complex by a bacterial catalytic scaffold. Nature, 469(7328), 107-113. https://doi.org/10.1038/nature09593

The assembly of a GTPase-kinase signalling complex by a bacterial catalytic scaffold. / Selyunin, Andrey S.; Sutton, Sarah E.; Weigele, Bethany A.; Reddick, L. Evan; Orchard, Robert C.; Bresson, Stefan M.; Tomchick, Diana R.; Alto, Neal M.

In: Nature, Vol. 469, No. 7328, 06.01.2011, p. 107-113.

Research output: Contribution to journalArticle

Selyunin, AS, Sutton, SE, Weigele, BA, Reddick, LE, Orchard, RC, Bresson, SM, Tomchick, DR & Alto, NM 2011, 'The assembly of a GTPase-kinase signalling complex by a bacterial catalytic scaffold', Nature, vol. 469, no. 7328, pp. 107-113. https://doi.org/10.1038/nature09593
Selyunin AS, Sutton SE, Weigele BA, Reddick LE, Orchard RC, Bresson SM et al. The assembly of a GTPase-kinase signalling complex by a bacterial catalytic scaffold. Nature. 2011 Jan 6;469(7328):107-113. https://doi.org/10.1038/nature09593
Selyunin, Andrey S. ; Sutton, Sarah E. ; Weigele, Bethany A. ; Reddick, L. Evan ; Orchard, Robert C. ; Bresson, Stefan M. ; Tomchick, Diana R. ; Alto, Neal M. / The assembly of a GTPase-kinase signalling complex by a bacterial catalytic scaffold. In: Nature. 2011 ; Vol. 469, No. 7328. pp. 107-113.
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