EspL is a bacterial cysteine protease effector that cleaves RHIM proteins to block necroptosis and inflammation

Jaclyn S. Pearson, Cristina Giogha, Sabrina Mühlen, Ueli Nachbur, Chi L L Pham, Ying Zhang, Joanne M Hildebrand, Clare V. Oates, Tania Wong Fok Lung, Danielle J. Ingle, Laura F. Dagley, Aleksandra Bankovacki, Emma J Petrie, Gunnar N Schroeder, Valerie F Crepin, Gad Frankel, Seth L. Masters, James Vince, James M. Murphy, Margaret Sunde & 3 others Andrew I. Webb, John Silke, Elizabeth L. Hartland

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Cell death signalling pathways contribute to tissue homeostasis and provide innate protection from infection. Adaptor proteins such as receptor-interacting serine/threonine-protein kinase 1 (RIPK1), receptor-interacting serine/threonine-protein kinase 3 (RIPK3), TIR-domain-containing adapter-inducing interferon-β (TRIF) and Z-DNA-binding protein 1 (ZBP1)/DNA-dependent activator of IFN-regulatory factors (DAI) that contain receptor-interacting protein (RIP) homotypic interaction motifs (RHIM) play a key role in cell death and inflammatory signalling 1-3. RHIM-dependent interactions help drive a caspase-independent form of cell death termed necroptosis 4,5. Here, we report that the bacterial pathogen enteropathogenic Escherichia coli (EPEC) uses the type III secretion system (T3SS) effector EspL to degrade the RHIM-containing proteins RIPK1, RIPK3, TRIF and ZBP1/DAI during infection. This requires a previously unrecognized tripartite cysteine protease motif in EspL (Cys47, His131, Asp153) that cleaves within the RHIM of these proteins. Bacterial infection and/or ectopic expression of EspL leads to rapid inactivation of RIPK1, RIPK3, TRIF and ZBP1/DAI and inhibition of tumour necrosis factor (TNF), lipopolysaccharide or polyinosinic:polycytidylic acid (poly(I:C))-induced necroptosis and inflammatory signalling. Furthermore, EPEC infection inhibits TNF-induced phosphorylation and plasma membrane localization of mixed lineage kinase domain-like pseudokinase (MLKL). In vivo, EspL cysteine protease activity contributes to persistent colonization of mice by the EPEC-like mouse pathogen Citrobacter rodentium. The activity of EspL defines a family of T3SS cysteine protease effectors found in a range of bacteria and reveals a mechanism by which gastrointestinal pathogens directly target RHIM-dependent inflammatory and necroptotic signalling pathways.

Original languageEnglish
Article number16258
Number of pages10
JournalNature Microbiology
Volume2
DOIs
Publication statusPublished - 13 Jan 2017
Externally publishedYes

Cite this

Pearson, Jaclyn S. ; Giogha, Cristina ; Mühlen, Sabrina ; Nachbur, Ueli ; Pham, Chi L L ; Zhang, Ying ; Hildebrand, Joanne M ; Oates, Clare V. ; Lung, Tania Wong Fok ; Ingle, Danielle J. ; Dagley, Laura F. ; Bankovacki, Aleksandra ; Petrie, Emma J ; Schroeder, Gunnar N ; Crepin, Valerie F ; Frankel, Gad ; Masters, Seth L. ; Vince, James ; Murphy, James M. ; Sunde, Margaret ; Webb, Andrew I. ; Silke, John ; Hartland, Elizabeth L. / EspL is a bacterial cysteine protease effector that cleaves RHIM proteins to block necroptosis and inflammation. In: Nature Microbiology. 2017 ; Vol. 2.
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title = "EspL is a bacterial cysteine protease effector that cleaves RHIM proteins to block necroptosis and inflammation",
abstract = "Cell death signalling pathways contribute to tissue homeostasis and provide innate protection from infection. Adaptor proteins such as receptor-interacting serine/threonine-protein kinase 1 (RIPK1), receptor-interacting serine/threonine-protein kinase 3 (RIPK3), TIR-domain-containing adapter-inducing interferon-β (TRIF) and Z-DNA-binding protein 1 (ZBP1)/DNA-dependent activator of IFN-regulatory factors (DAI) that contain receptor-interacting protein (RIP) homotypic interaction motifs (RHIM) play a key role in cell death and inflammatory signalling 1-3. RHIM-dependent interactions help drive a caspase-independent form of cell death termed necroptosis 4,5. Here, we report that the bacterial pathogen enteropathogenic Escherichia coli (EPEC) uses the type III secretion system (T3SS) effector EspL to degrade the RHIM-containing proteins RIPK1, RIPK3, TRIF and ZBP1/DAI during infection. This requires a previously unrecognized tripartite cysteine protease motif in EspL (Cys47, His131, Asp153) that cleaves within the RHIM of these proteins. Bacterial infection and/or ectopic expression of EspL leads to rapid inactivation of RIPK1, RIPK3, TRIF and ZBP1/DAI and inhibition of tumour necrosis factor (TNF), lipopolysaccharide or polyinosinic:polycytidylic acid (poly(I:C))-induced necroptosis and inflammatory signalling. Furthermore, EPEC infection inhibits TNF-induced phosphorylation and plasma membrane localization of mixed lineage kinase domain-like pseudokinase (MLKL). In vivo, EspL cysteine protease activity contributes to persistent colonization of mice by the EPEC-like mouse pathogen Citrobacter rodentium. The activity of EspL defines a family of T3SS cysteine protease effectors found in a range of bacteria and reveals a mechanism by which gastrointestinal pathogens directly target RHIM-dependent inflammatory and necroptotic signalling pathways.",
author = "Pearson, {Jaclyn S.} and Cristina Giogha and Sabrina M{\"u}hlen and Ueli Nachbur and Pham, {Chi L L} and Ying Zhang and Hildebrand, {Joanne M} and Oates, {Clare V.} and Lung, {Tania Wong Fok} and Ingle, {Danielle J.} and Dagley, {Laura F.} and Aleksandra Bankovacki and Petrie, {Emma J} and Schroeder, {Gunnar N} and Crepin, {Valerie F} and Gad Frankel and Masters, {Seth L.} and James Vince and Murphy, {James M.} and Margaret Sunde and Webb, {Andrew I.} and John Silke and Hartland, {Elizabeth L.}",
year = "2017",
month = "1",
day = "13",
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Pearson, JS, Giogha, C, Mühlen, S, Nachbur, U, Pham, CLL, Zhang, Y, Hildebrand, JM, Oates, CV, Lung, TWF, Ingle, DJ, Dagley, LF, Bankovacki, A, Petrie, EJ, Schroeder, GN, Crepin, VF, Frankel, G, Masters, SL, Vince, J, Murphy, JM, Sunde, M, Webb, AI, Silke, J & Hartland, EL 2017, 'EspL is a bacterial cysteine protease effector that cleaves RHIM proteins to block necroptosis and inflammation', Nature Microbiology, vol. 2, 16258. https://doi.org/10.1038/nmicrobiol.2016.258

EspL is a bacterial cysteine protease effector that cleaves RHIM proteins to block necroptosis and inflammation. / Pearson, Jaclyn S.; Giogha, Cristina; Mühlen, Sabrina; Nachbur, Ueli; Pham, Chi L L; Zhang, Ying; Hildebrand, Joanne M; Oates, Clare V.; Lung, Tania Wong Fok; Ingle, Danielle J.; Dagley, Laura F.; Bankovacki, Aleksandra; Petrie, Emma J; Schroeder, Gunnar N; Crepin, Valerie F; Frankel, Gad; Masters, Seth L.; Vince, James ; Murphy, James M.; Sunde, Margaret; Webb, Andrew I.; Silke, John; Hartland, Elizabeth L.

In: Nature Microbiology, Vol. 2, 16258, 13.01.2017.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - EspL is a bacterial cysteine protease effector that cleaves RHIM proteins to block necroptosis and inflammation

AU - Pearson, Jaclyn S.

AU - Giogha, Cristina

AU - Mühlen, Sabrina

AU - Nachbur, Ueli

AU - Pham, Chi L L

AU - Zhang, Ying

AU - Hildebrand, Joanne M

AU - Oates, Clare V.

AU - Lung, Tania Wong Fok

AU - Ingle, Danielle J.

AU - Dagley, Laura F.

AU - Bankovacki, Aleksandra

AU - Petrie, Emma J

AU - Schroeder, Gunnar N

AU - Crepin, Valerie F

AU - Frankel, Gad

AU - Masters, Seth L.

AU - Vince, James

AU - Murphy, James M.

AU - Sunde, Margaret

AU - Webb, Andrew I.

AU - Silke, John

AU - Hartland, Elizabeth L.

PY - 2017/1/13

Y1 - 2017/1/13

N2 - Cell death signalling pathways contribute to tissue homeostasis and provide innate protection from infection. Adaptor proteins such as receptor-interacting serine/threonine-protein kinase 1 (RIPK1), receptor-interacting serine/threonine-protein kinase 3 (RIPK3), TIR-domain-containing adapter-inducing interferon-β (TRIF) and Z-DNA-binding protein 1 (ZBP1)/DNA-dependent activator of IFN-regulatory factors (DAI) that contain receptor-interacting protein (RIP) homotypic interaction motifs (RHIM) play a key role in cell death and inflammatory signalling 1-3. RHIM-dependent interactions help drive a caspase-independent form of cell death termed necroptosis 4,5. Here, we report that the bacterial pathogen enteropathogenic Escherichia coli (EPEC) uses the type III secretion system (T3SS) effector EspL to degrade the RHIM-containing proteins RIPK1, RIPK3, TRIF and ZBP1/DAI during infection. This requires a previously unrecognized tripartite cysteine protease motif in EspL (Cys47, His131, Asp153) that cleaves within the RHIM of these proteins. Bacterial infection and/or ectopic expression of EspL leads to rapid inactivation of RIPK1, RIPK3, TRIF and ZBP1/DAI and inhibition of tumour necrosis factor (TNF), lipopolysaccharide or polyinosinic:polycytidylic acid (poly(I:C))-induced necroptosis and inflammatory signalling. Furthermore, EPEC infection inhibits TNF-induced phosphorylation and plasma membrane localization of mixed lineage kinase domain-like pseudokinase (MLKL). In vivo, EspL cysteine protease activity contributes to persistent colonization of mice by the EPEC-like mouse pathogen Citrobacter rodentium. The activity of EspL defines a family of T3SS cysteine protease effectors found in a range of bacteria and reveals a mechanism by which gastrointestinal pathogens directly target RHIM-dependent inflammatory and necroptotic signalling pathways.

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