Inhibitory mechanism of Escherichia coli RelE-RelB toxin-antitoxin module involves a helix displacement near an mRNA interferase active site.
In Escherichia coli, RelE toxin participates in growth arrest and cell death by inducing mRNA degradation at the ribosomal A-site under stress conditions. The NMR structures of a mutant of E. coli RelE toxin, RelE(R81A/R83A), with reduced toxicity and its complex with an inhibitory peptide from RelB antitoxin, RelB(C) (Lys(47)-Leu(79)), have been determined. In the free RelE(R81A/R83A) structure, helix alpha4 at the C terminus adopts a closed conformation contacting with the beta-sheet core and adjacent loops. In the RelE(R81A/R83A)-RelB(C) complex, helix alpha3(*) of RelB(C) displaces alpha4 of RelE(R81A/R83A) from the binding site on the beta-sheet core. This helix replacement results in neutralization of a conserved positively charged cluster of RelE by acidic residues from alpha3(*) of RelB. The released helix alpha4 becomes unfolded, adopting an open conformation with increased mobility. The displacement of alpha4 disrupts the geometry of critical residues, including Arg(81) and Tyr(87), in a putative active site of RelE toxin. Our structures indicate that RelB counteracts the toxic activity of RelE by displacing alpha4 helix from the catalytically competent position found in the free RelE structure.
Li
Guang-Yao
GY
Division of Signaling Biology, Ontario Cancer Institute, Toronto, Ontario, Canada.
Zhang
Yonglong
Y
Inouye
Masayori
M
Ikura
Mitsuhiko
M
D019943:Amino Acid Substitution; D000992:Antitoxins / Q000737:chemistry*; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D055162:Biocatalysis; D020134:Catalytic Domain; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D011485:Protein Binding; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000737:chemistry*; D014118:Toxins, Biological / Q000737:chemistry*; D051841:Transcription Factor RelB
D000992:Antitoxins; D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D012333:RNA, Messenger; D014118:Toxins, Biological; C465567:RelB protein, E coli; D051841:Transcription Factor RelB; C465566:RelE protein, E coli