Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin-antitoxin system
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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.
In: eLife, Vol. 12, RP90400, 2023.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Structural basis for kinase inhibition in the tripartite E. coli HipBST toxin-antitoxin system
AU - Bærentsen, René L.
AU - Nielsen, Stine V.
AU - Skjerning, Ragnhild B.
AU - Lyngsø, Jeppe
AU - Bisiak, Francesco
AU - Pedersen, Jan Skov
AU - Gerdes, Kenn
AU - Sørensen, Michael A.
AU - Brodersen, Ditlev E.
N1 - Publisher Copyright: © 2023, Bærentsen, Nielsen et al.
PY - 2023
Y1 - 2023
N2 - 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.
AB - 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.
KW - E. coli
KW - HipBA
KW - infectious disease
KW - microbiology
KW - molecular biophysics
KW - Ser/Thr kinase
KW - STK
KW - structural biology
KW - tRNA synthetase
KW - TrpS
U2 - 10.7554/eLife.90400
DO - 10.7554/eLife.90400
M3 - Journal article
C2 - 37929938
AN - SCOPUS:85176293146
VL - 12
JO - eLife
JF - eLife
SN - 2050-084X
M1 - RP90400
ER -
ID: 373659915