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Crystal structure of bacterial cytotoxic necrotizing factor CNFy reveals molecular building blocks for intoxication.

Chaoprasid, Paweena
; 
Lukat, Peer
; 
Mühlen, Sabrina
; 
Heidler, Thomas
; 
Gazdag, Emerich-Mihai
; 
Dong, Shuangshuang
; 
Bi, Wenjie
; 
Rüter, Christian
; 
Kirchenwitz, Marco
; 
Steffen, Anika
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2021-01-07
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Abstract
Cytotoxic necrotizing factors (CNFs) are bacterial single-chain exotoxins that modulate cytokinetic/oncogenic and inflammatory processes through activation of host cell Rho GTPases. To achieve this, they are secreted, bind surface receptors to induce endocytosis and translocate a catalytic unit into the cytosol to intoxicate host cells. A three-dimensional structure that provides insight into the underlying mechanisms is still lacking. Here, we determined the crystal structure of full-length Yersinia pseudotuberculosis CNFY . CNFY consists of five domains (D1-D5), and by integrating structural and functional data, we demonstrate that D1-3 act as export and translocation module for the catalytic unit (D4-5) and for a fused β-lactamase reporter protein. We further found that D4, which possesses structural similarity to ADP-ribosyl transferases, but had no equivalent catalytic activity, changed its position to interact extensively with D5 in the crystal structure of the free D4-5 fragment. This liberates D5 from a semi-blocked conformation in full-length CNFY , leading to higher deamidation activity. Finally, we identify CNF translocation modules in several uncharacterized fusion proteins, which suggests their usability as a broad-specificity protein delivery tool.
Citation
EMBO J. 2021 Jan 7:e105202. doi: 10.15252/embj.2020105202. Epub ahead of print.
Affiliation
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
Publisher
Springer
Journal
The EMBO journal
URI
http://hdl.handle.net/10033/622742
DOI
10.15252/embj.2020105202
PubMed ID
33410511
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en
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1460-2075
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Attribution 4.0 International
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Publications of the research group structure and functions of Proteins(SFPR)
publications of the research group cell biology (ZBIO)
publications of the research group cellular proteom research (CPRO)
publications of the scientific administration (GFW)