Binding specificity of type three secretion system effector NleH2 to multi-cargo chaperone CesT and their phosphorylation.

Publisher: Cold Spring Harbor Laboratory Press Country of Publication: United States NLM ID: 9211750 Publication Model: Print Cited Medium: Internet ISSN: 1469-896X (Electronic) Linking ISSN: 09618368 NLM ISO Abbreviation: Protein Sci Subsets: MEDLINE

Publication: 2001- : Woodbury, NY : Cold Spring Harbor Laboratory Press
Original Publication: New York, N.Y. : Cambridge University Press, c1992-

Enteropathogenic Escherichia coli/*genetics ; Escherichia coli Proteins/*chemistry ; Molecular Chaperones/*chemistry ; RNA-Binding Proteins/*chemistry ; Receptors, Cell Surface/*chemistry ; Repressor Proteins/*chemistry; Amino Acid Sequence ; Binding Sites ; Cloning, Molecular ; Enteropathogenic Escherichia coli/metabolism ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; Gene Expression ; Gene Expression Regulation, Bacterial ; Genetic Vectors/chemistry ; Genetic Vectors/metabolism ; Kinetics ; Models, Molecular ; Molecular Chaperones/genetics ; Molecular Chaperones/metabolism ; Phosphorylation ; Protein Binding ; Protein Conformation, alpha-Helical ; Protein Conformation, beta-Strand ; Protein Interaction Domains and Motifs ; Protein Processing, Post-Translational ; RNA-Binding Proteins/genetics ; RNA-Binding Proteins/metabolism ; Receptors, Cell Surface/genetics ; Receptors, Cell Surface/metabolism ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Repressor Proteins/genetics ; Repressor Proteins/metabolism ; Sequence Alignment ; Sequence Homology, Amino Acid ; Substrate Specificity ; Type III Secretion Systems/genetics ; Type III Secretion Systems/metabolism

Gram-negative pathogens like Enteropathogenic Escherichia coli (EPEC) utilize the type three secretion system (T3SS) to translocate various effector proteins that are needed to "hijack" the host system for pathogenic survival. Specialized T3SS chaperones inside bacterial cells stabilize these effector proteins and facilitate their translocation. CesT is a unique multi-cargo chaperone that interacts with and translocates ~10 different effector proteins. Here, we report the specific interaction between CesT and its key effector, NleH2, and explore the potential role of NleH2 as a kinase for CesT phosphorylation. First, we identified the chaperone-binding domain (CBD; 19-97aa) of NleH2, and mapped the specific interaction sites for both CesT and NleH2. The N- and C-terminal residues of the CBD interact with the dimeric interface of CesT. Further, we compared the CesT binding to NleH2, to that of another key effector Tir and with the global carbon regulator CsrA. Notably, the effectors have the binding regions at the β-sheet core and dimer interface of CesT, whereas the CsrA regulator interacts predominantly through the C-terminal region, which is found ~17 Å away from the effectors-binding sites. Next, we showed that NleH2 remains an active kinase even as a complex with CesT and is responsible for its autophosphorylation as well as phosphorylation of CesT at Tyr153. Collectively, our findings enhance the understanding of the role of multi-cargo chaperone CesT in orchestrating effector translocation through T3SS.
(© 2021 The Protein Society.)

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Keywords: chaperone; effectors; kinase; pathogen; phosphorylation; secretion system

0 (CesT protein, E coli)
0 (CsrA protein, E coli)
0 (Escherichia coli Proteins)
0 (Molecular Chaperones)
0 (NleH protein, E coli)
0 (RNA-Binding Proteins)
0 (Receptors, Cell Surface)
0 (Recombinant Proteins)
0 (Repressor Proteins)
0 (Tir protein, E coli)
0 (Type III Secretion Systems)

Date Created: 20211018 Date Completed: 20220208 Latest Revision: 20221202

20221202

PMC8605367

10.1002/pro.4210

34662450