Bacterial factors drive the differential targeting of Guanylate Binding Proteins to<i>Francisella</i>and<i>Shigella</i>
ABSTRACT Guanylate-Binding Proteins (GBPs) are interferon-inducible GTPases that play a key role in cell autonomous responses against intracellular pathogens. Seven GBPs are present in humans. Despite sharing high sequence similarity, subtle differences among GBPs translate into functional divergences that are still largely not understood. A key step for the antimicrobial activity of GBPs towards cytosolic bacteria is the formation of supramolecular GBP complexes on the bacterial surface. Such complexes are formed when GBP1 binds lipopolysaccharide (LPS) fromShigellaandSalmonellaand further recruits GBP2, 3, and 4.Here, we investigated GBPs recruitment onFrancisella novicida, a professional cytosol-dwelling pathogen with an atypical tetra-acylated LPS. Co-infection experiments demonstrated that GBPs target preferentiallyS. flexnericompared toF. novicida.F. novicidawas coated by GBP1 and GBP2 in human macrophages but escaped targeting by GBP3 and GBP4. GBP1 and GBP2 features that drive recruitment toF. novicidawere investigated revealing that GBP1 GDPase activity is required to initiate GBP recruitment toF. novicidabut facultative to targetS. flexneri. Furthermore, analysis of chimeric GBP2/5 proteins identified a central domain in GBP2 necessary and sufficient to targetF. novicida.Finally, aF. novicidaΔlpxFmutant with a penta-acylated lipid A was targeted by GBP3 suggesting that lipid A tetra-acylation contributes to escape from GBP3. Altogether our results indicate that GBPs have different affinity for different bacteria and that the repertoire of GBPs recruited onto cytosolic bacteria is dictated by GBP-intrinsic features and specific bacterial factors, including the structure of the lipid A.IMPORTANCE Few bacteria have adapted to thrive in the hostile environment of the cell cytosol. As a professional cytosol-dwelling pathogen,S. flexnerisecretes several effectors to block cytosolic immune effectors, including GBPs. This study illustrates a different approach of adapting to the host cytosol: the stealth strategy developed byF. novicida.F. novicidabears an atypical hypoacylated LPS, which does not elicit neither TLR4 nor caspase-11 activation. Here, this atypical LPS was shown to promote escape from GBP3 targeting. Furthermore, the lower affinity of GBPs forF. novicidaallowed to decipher the different domains that govern GBP recruitment to the bacterial surface. This study illustrates the importance of investigating different bacterial models to broaden our understanding of the intricacies of host-pathogen interactions..
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Preprint| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
bioRxiv.org - (2024) vom: 23. Apr. Zur Gesamtaufnahme - year:2024 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Valeva, Stanimira V. [VerfasserIn] |
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| doi: |
10.1101/2021.06.16.448779 |
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| PPN (Katalog-ID): |
XBI03201600X |
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| 245 | 1 | 0 | |a Bacterial factors drive the differential targeting of Guanylate Binding Proteins to<i>Francisella</i>and<i>Shigella</i> |
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| 520 | |a ABSTRACT Guanylate-Binding Proteins (GBPs) are interferon-inducible GTPases that play a key role in cell autonomous responses against intracellular pathogens. Seven GBPs are present in humans. Despite sharing high sequence similarity, subtle differences among GBPs translate into functional divergences that are still largely not understood. A key step for the antimicrobial activity of GBPs towards cytosolic bacteria is the formation of supramolecular GBP complexes on the bacterial surface. Such complexes are formed when GBP1 binds lipopolysaccharide (LPS) fromShigellaandSalmonellaand further recruits GBP2, 3, and 4.Here, we investigated GBPs recruitment onFrancisella novicida, a professional cytosol-dwelling pathogen with an atypical tetra-acylated LPS. Co-infection experiments demonstrated that GBPs target preferentiallyS. flexnericompared toF. novicida.F. novicidawas coated by GBP1 and GBP2 in human macrophages but escaped targeting by GBP3 and GBP4. GBP1 and GBP2 features that drive recruitment toF. novicidawere investigated revealing that GBP1 GDPase activity is required to initiate GBP recruitment toF. novicidabut facultative to targetS. flexneri. Furthermore, analysis of chimeric GBP2/5 proteins identified a central domain in GBP2 necessary and sufficient to targetF. novicida.Finally, aF. novicidaΔlpxFmutant with a penta-acylated lipid A was targeted by GBP3 suggesting that lipid A tetra-acylation contributes to escape from GBP3. Altogether our results indicate that GBPs have different affinity for different bacteria and that the repertoire of GBPs recruited onto cytosolic bacteria is dictated by GBP-intrinsic features and specific bacterial factors, including the structure of the lipid A.IMPORTANCE Few bacteria have adapted to thrive in the hostile environment of the cell cytosol. As a professional cytosol-dwelling pathogen,S. flexnerisecretes several effectors to block cytosolic immune effectors, including GBPs. This study illustrates a different approach of adapting to the host cytosol: the stealth strategy developed byF. novicida.F. novicidabears an atypical hypoacylated LPS, which does not elicit neither TLR4 nor caspase-11 activation. Here, this atypical LPS was shown to promote escape from GBP3 targeting. Furthermore, the lower affinity of GBPs forF. novicidaallowed to decipher the different domains that govern GBP recruitment to the bacterial surface. This study illustrates the importance of investigating different bacterial models to broaden our understanding of the intricacies of host-pathogen interactions. | ||
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| 700 | 1 | |a Lagrange, Brice |e verfasserin |4 aut | |
| 700 | 1 | |a Henry, Thomas |e verfasserin |4 aut | |
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