Mechanisms of Yersinia YopO kinase substrate specificity

Wei Lin Lee, Pavithra Singaravelu, Sheena Wee, Bo Xue, Khay Chun Ang, Jayantha Gunaratne, Jonathan M. Grimes, Kunchithapadam Swaminathan, Robert C. Robinson

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Yersinia bacteria cause a range of human diseases, including yersiniosis, Far East scarlet-like fever and the plague. Yersiniae modulate and evade host immune defences through injection of Yersinia outer proteins (Yops) into phagocytic cells. One of the Yops, YopO (also known as YpkA) obstructs phagocytosis through disrupting actin filament regulation processes - inhibiting polymerization-promoting signaling through sequestration of Rac/Rho family GTPases and by using monomeric actin as bait to recruit and phosphorylate host actin-regulating proteins. Here we set out to identify mechanisms of specificity in protein phosphorylation by YopO that would clarify its effects on cytoskeleton disruption. We report the MgADP structure of Yersinia enterocolitica YopO in complex with actin, which reveals its active site architecture. Using a proteome-wide kinase-interacting substrate screening (KISS) method, we identified that YopO phosphorylates a wide range of actin-modulating proteins and located their phosphorylation sites by mass spectrometry. Using artificial substrates we clarified YopO's substrate length requirements and its phosphorylation consensus sequence. These findings provide fresh insight into the mechanism of the YopO kinase and demonstrate that YopO executes a specific strategy targeting actin-modulating proteins, across multiple functionalities, to compete for control of their native phospho-signaling, thus hampering the cytoskeletal processes required for macrophage phagocytosis.

Original languageEnglish
Article number39998
JournalScientific reports
Volume7
DOIs
Publication statusPublished - Jan 4 2017
Externally publishedYes

Fingerprint

Yersinia
Substrate Specificity
Phosphotransferases
Actins
Phosphorylation
Phagocytosis
Proteins
Scarlet Fever
rho GTP-Binding Proteins
Yersinia enterocolitica
Plague
Far East
Consensus Sequence
Proteome
Phagocytes
Cytoskeleton
Actin Cytoskeleton
Polymerization
Adenosine Diphosphate
Mass Spectrometry

ASJC Scopus subject areas

  • General

Cite this

Lee, W. L., Singaravelu, P., Wee, S., Xue, B., Ang, K. C., Gunaratne, J., ... Robinson, R. C. (2017). Mechanisms of Yersinia YopO kinase substrate specificity. Scientific reports, 7, [39998]. https://doi.org/10.1038/srep39998

Mechanisms of Yersinia YopO kinase substrate specificity. / Lee, Wei Lin; Singaravelu, Pavithra; Wee, Sheena; Xue, Bo; Ang, Khay Chun; Gunaratne, Jayantha; Grimes, Jonathan M.; Swaminathan, Kunchithapadam; Robinson, Robert C.

In: Scientific reports, Vol. 7, 39998, 04.01.2017.

Research output: Contribution to journalArticle

Lee, WL, Singaravelu, P, Wee, S, Xue, B, Ang, KC, Gunaratne, J, Grimes, JM, Swaminathan, K & Robinson, RC 2017, 'Mechanisms of Yersinia YopO kinase substrate specificity', Scientific reports, vol. 7, 39998. https://doi.org/10.1038/srep39998
Lee WL, Singaravelu P, Wee S, Xue B, Ang KC, Gunaratne J et al. Mechanisms of Yersinia YopO kinase substrate specificity. Scientific reports. 2017 Jan 4;7. 39998. https://doi.org/10.1038/srep39998
Lee, Wei Lin ; Singaravelu, Pavithra ; Wee, Sheena ; Xue, Bo ; Ang, Khay Chun ; Gunaratne, Jayantha ; Grimes, Jonathan M. ; Swaminathan, Kunchithapadam ; Robinson, Robert C. / Mechanisms of Yersinia YopO kinase substrate specificity. In: Scientific reports. 2017 ; Vol. 7.
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