Structure of the Shigella T3SS effector IpaH defines a new class of E3 ubiquitin ligases

Alexander U. Singer; John R. Rohde; Robert Lam; Tatiana Skarina; Olga Kagan; Rosa DiLeo; Nickolay Y. Chirgadze; Marianne E. Cuff; Andrzej Joachimiak; Mike Tyers; Philippe J. Sansonetti; Claude Parsot; Alexei Savchenko

doi:10.1038/nsmb.1511

Structure of the Shigella T3SS effector IpaH defines a new class of E3 ubiquitin ligases

Alexander U. Singer, John R. Rohde, Robert Lam, Tatiana Skarina, Olga Kagan, Rosa DiLeo, Nickolay Y. Chirgadze, Marianne E. Cuff, Andrzej Joachimiak, Mike Tyers, Philippe J. Sansonetti, Claude Parsot, Alexei Savchenko

Research output: Contribution to journal › Article › peer-review

135 Citations (Scopus)

Abstract

IpaH proteins are E3 ubiquitin ligases delivered by the type III secretion apparatus into host cells upon infection of humans by the Gram-negative pathogen Shigella flexneri. These proteins comprise a variable leucine-rich repeat-containing N-terminal domain and a conserved C-terminal domain harboring an invariant cysteine residue that is crucial for activity. IpaH homologs are encoded by diverse animal and plant pathogens. Here we demonstrate that the IpaH C-terminal domain carries the catalytic activity for ubiquitin transfer and that the N-terminal domain carries the substrate specificity. The structure of the IpaH C-terminal domain, determined to 2.65-Å resolution, represents an all-helical fold bearing no resemblance to previously defined E3 ubiquitin ligases. The conserved and essential cysteine residue lies on a flexible, surface-exposed loop surrounded by conserved acidic residues, two of which are crucial for IpaH activity.

Original language	English
Pages (from-to)	1293-1301
Number of pages	9
Journal	Nature Structural and Molecular Biology
Volume	15
Issue number	12
DOIs	https://doi.org/10.1038/nsmb.1511
Publication status	Published - Dec 2008
Externally published	Yes

Bibliographical note

Funding Information:
We wish to thank the staff at the Argonne National Laboratory beam line 19-ID for assistance with data collection and A. Edwards for critical reading of the manuscript. We also wish to thank S. Dhe-Paganon and G. Avvakumov at the Structural Genomics Consortium, Toronto, for providing the collection of expression constructs for human E2-conjugating enzymes. We thank D. Briant for insight to E3 ubiquitination assays. This work was supported by US National Institutes of Health Grants GM62414-01, by the Ontario Research and Development Challenge Fund and by a grant from the Canadian Institutes of Health Research Grant. M.T. is supported by grants from the Canadian Institutes of Health Research (MT012466 and MOP-57795), the National Cancer Institute of Canada, the Royal Society and the Scottish Universities Life Sciences Alliance.

ASJC Scopus Subject Areas

Structural Biology
Molecular Biology

PubMed: MeSH publication types

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

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Cite this

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title = "Structure of the Shigella T3SS effector IpaH defines a new class of E3 ubiquitin ligases",

abstract = "IpaH proteins are E3 ubiquitin ligases delivered by the type III secretion apparatus into host cells upon infection of humans by the Gram-negative pathogen Shigella flexneri. These proteins comprise a variable leucine-rich repeat-containing N-terminal domain and a conserved C-terminal domain harboring an invariant cysteine residue that is crucial for activity. IpaH homologs are encoded by diverse animal and plant pathogens. Here we demonstrate that the IpaH C-terminal domain carries the catalytic activity for ubiquitin transfer and that the N-terminal domain carries the substrate specificity. The structure of the IpaH C-terminal domain, determined to 2.65-{\AA} resolution, represents an all-helical fold bearing no resemblance to previously defined E3 ubiquitin ligases. The conserved and essential cysteine residue lies on a flexible, surface-exposed loop surrounded by conserved acidic residues, two of which are crucial for IpaH activity.",

author = "Singer, {Alexander U.} and Rohde, {John R.} and Robert Lam and Tatiana Skarina and Olga Kagan and Rosa DiLeo and Chirgadze, {Nickolay Y.} and Cuff, {Marianne E.} and Andrzej Joachimiak and Mike Tyers and Sansonetti, {Philippe J.} and Claude Parsot and Alexei Savchenko",

note = "Funding Information: We wish to thank the staff at the Argonne National Laboratory beam line 19-ID for assistance with data collection and A. Edwards for critical reading of the manuscript. We also wish to thank S. Dhe-Paganon and G. Avvakumov at the Structural Genomics Consortium, Toronto, for providing the collection of expression constructs for human E2-conjugating enzymes. We thank D. Briant for insight to E3 ubiquitination assays. This work was supported by US National Institutes of Health Grants GM62414-01, by the Ontario Research and Development Challenge Fund and by a grant from the Canadian Institutes of Health Research Grant. M.T. is supported by grants from the Canadian Institutes of Health Research (MT012466 and MOP-57795), the National Cancer Institute of Canada, the Royal Society and the Scottish Universities Life Sciences Alliance.",

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AU - DiLeo, Rosa

AU - Chirgadze, Nickolay Y.

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AU - Joachimiak, Andrzej

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AU - Sansonetti, Philippe J.

AU - Parsot, Claude

AU - Savchenko, Alexei

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Structure of the Shigella T3SS effector IpaH defines a new class of E3 ubiquitin ligases

Abstract

Bibliographical note

ASJC Scopus Subject Areas

PubMed: MeSH publication types

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