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TitleMechanism of threonine ADP-ribosylation of F-actin by a Tc toxin.
Journal, issue, pagesNat Commun, Vol. 13, Issue 1, Page 4202, Year 2022
Publish dateJul 20, 2022
AuthorsAlexander Belyy / Florian Lindemann / Daniel Roderer / Johanna Funk / Benjamin Bardiaux / Jonas Protze / Peter Bieling / Hartmut Oschkinat / Stefan Raunser /
PubMed AbstractTc toxins deliver toxic enzymes into host cells by a unique injection mechanism. One of these enzymes is the actin ADP-ribosyltransferase TccC3, whose activity leads to the clustering of the cellular ...Tc toxins deliver toxic enzymes into host cells by a unique injection mechanism. One of these enzymes is the actin ADP-ribosyltransferase TccC3, whose activity leads to the clustering of the cellular cytoskeleton and ultimately cell death. Here, we show in atomic detail how TccC3 modifies actin. We find that the ADP-ribosyltransferase does not bind to G-actin but interacts with two consecutive actin subunits of F-actin. The binding of TccC3 to F-actin occurs via an induced-fit mechanism that facilitates access of NAD to the nucleotide binding pocket. The following nucleophilic substitution reaction results in the transfer of ADP-ribose to threonine-148 of F-actin. We demonstrate that this site-specific modification of F-actin prevents its interaction with depolymerization factors, such as cofilin, which impairs actin network turnover and leads to steady actin polymerization. Our findings reveal in atomic detail a mechanism of action of a bacterial toxin through specific targeting and modification of F-actin.
External linksNat Commun / PubMed:35858890 / PubMed Central
MethodsEM (single particle) / NMR (solution)
Resolution3.5 - 3.8 Å
Structure data

14532

7z7h

EMDB-14532, PDB-7z7h:
Structure of P. luminescens TccC3-F-actin complex
Method: EM (single particle) / Resolution: 3.8 Å

14533

7z7i

EMDB-14533, PDB-7z7i:
Structure of ADP-ribosylated F-actin
Method: EM (single particle) / Resolution: 3.5 Å

PDB-7zbq:
Structure of the ADP-ribosyltransferase TccC3HVR from Photorhabdus Luminescens
Method: SOLUTION NMR

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

ChemComp-MG:
Unknown entry

ChemComp-APR:
ADENOSINE-5-DIPHOSPHORIBOSE

ChemComp-NCA:
NICOTINAMIDE / medication*YM

Source
  • photorhabdus luminescens (bacteria)
  • rabbit (rabbit)
  • oryctolagus cuniculus (rabbit)
KeywordsTOXIN / Bacterial toxin / F-actin / CYTOSOLIC PROTEIN / ADP-ribosylation / R-S-E motif / actin modification

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