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Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin–antitoxin system

Many bacteria encode multiple toxin–antitoxin (TA) systems targeting separate, but closely related, cellular functions. The toxin of the Escherichia coli hipBA system, HipA, is a kinase that inhibits translation via phosphorylation of glutamyl-tRNA synthetase. Enteropathogenic E. coli O127:H6 encode...

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Autores principales: Bærentsen, René L, Nielsen, Stine V, Skjerning, Ragnhild B, Lyngsø, Jeppe, Bisiak, Francesco, Pedersen, Jan Skov, Gerdes, Kenn, Sørensen, Michael A, Brodersen, Ditlev E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10627512/
https://www.ncbi.nlm.nih.gov/pubmed/37929938
http://dx.doi.org/10.7554/eLife.90400
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author Bærentsen, René L
Nielsen, Stine V
Skjerning, Ragnhild B
Lyngsø, Jeppe
Bisiak, Francesco
Pedersen, Jan Skov
Gerdes, Kenn
Sørensen, Michael A
Brodersen, Ditlev E
author_facet Bærentsen, René L
Nielsen, Stine V
Skjerning, Ragnhild B
Lyngsø, Jeppe
Bisiak, Francesco
Pedersen, Jan Skov
Gerdes, Kenn
Sørensen, Michael A
Brodersen, Ditlev E
author_sort Bærentsen, René L
collection PubMed
description Many bacteria encode multiple toxin–antitoxin (TA) systems targeting separate, but closely related, cellular functions. The toxin of the Escherichia coli hipBA system, HipA, is a kinase that inhibits translation via phosphorylation of glutamyl-tRNA synthetase. Enteropathogenic E. coli O127:H6 encodes the hipBA-like, tripartite TA system; hipBST, in which the HipT toxin specifically targets the tryptophanyl-tRNA synthetase, TrpS. Notably, in the tripartite system, the function as antitoxin has been taken over by the third protein, HipS, but the molecular details of how activity of HipT is inhibited remain poorly understood. Here, we show that HipBST is structurally different from E. coli HipBA and that the unique HipS protein, which is homologous to the N-terminal subdomain of HipA, inhibits the kinase through insertion of a conserved Trp residue into the active site. We also show how auto-phosphorylation at two conserved sites in the kinase toxin serve different roles and affect the ability of HipS to neutralize HipT. Finally, solution structural studies show how phosphorylation affects overall TA complex flexibility.
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spelling pubmed-106275122023-11-07 Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin–antitoxin system Bærentsen, René L Nielsen, Stine V Skjerning, Ragnhild B Lyngsø, Jeppe Bisiak, Francesco Pedersen, Jan Skov Gerdes, Kenn Sørensen, Michael A Brodersen, Ditlev E eLife Microbiology and Infectious Disease Many bacteria encode multiple toxin–antitoxin (TA) systems targeting separate, but closely related, cellular functions. The toxin of the Escherichia coli hipBA system, HipA, is a kinase that inhibits translation via phosphorylation of glutamyl-tRNA synthetase. Enteropathogenic E. coli O127:H6 encodes the hipBA-like, tripartite TA system; hipBST, in which the HipT toxin specifically targets the tryptophanyl-tRNA synthetase, TrpS. Notably, in the tripartite system, the function as antitoxin has been taken over by the third protein, HipS, but the molecular details of how activity of HipT is inhibited remain poorly understood. Here, we show that HipBST is structurally different from E. coli HipBA and that the unique HipS protein, which is homologous to the N-terminal subdomain of HipA, inhibits the kinase through insertion of a conserved Trp residue into the active site. We also show how auto-phosphorylation at two conserved sites in the kinase toxin serve different roles and affect the ability of HipS to neutralize HipT. Finally, solution structural studies show how phosphorylation affects overall TA complex flexibility. eLife Sciences Publications, Ltd 2023-11-06 /pmc/articles/PMC10627512/ /pubmed/37929938 http://dx.doi.org/10.7554/eLife.90400 Text en © 2023, Bærentsen, Nielsen et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Microbiology and Infectious Disease
Bærentsen, René L
Nielsen, Stine V
Skjerning, Ragnhild B
Lyngsø, Jeppe
Bisiak, Francesco
Pedersen, Jan Skov
Gerdes, Kenn
Sørensen, Michael A
Brodersen, Ditlev E
Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin–antitoxin system
title Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin–antitoxin system
title_full Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin–antitoxin system
title_fullStr Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin–antitoxin system
title_full_unstemmed Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin–antitoxin system
title_short Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin–antitoxin system
title_sort structural basis for kinase inhibition in the tripartite e. coli hipbst toxin–antitoxin system
topic Microbiology and Infectious Disease
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10627512/
https://www.ncbi.nlm.nih.gov/pubmed/37929938
http://dx.doi.org/10.7554/eLife.90400
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