Functional dissection of the retrograde Shiga toxin trafficking inhibitor Retro-2

Forrester, Alison; Rathjen, Stefan J.; García-Castillo, María; Bachert, Collin; Couhert, Audrey; Tepshi, Livia; Pichard, Sylvain; Martinez, Jennifer; Munier, Mathilde; Sierocki, Raphaël; Renard, Henri-François; Valades-Cruz, César Augusto; Dingli, Florent; Loew, Damarys; Lamaze, Christophe; Cintrat, Jean‐Christophe; Linstedt, Adam D.; Gillet, Daniel; Barbier, Julien; Johannes, Ludger

doi:10.1038/s41589-020-0474-4

Cited by 44 publications

(58 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This effect is rescued by expression of full-length GPP130, but not by GPP130 mutants that lack the binding site for STxB/STx1B or that cannot cycle between the Golgi and early endosomes [19]. Furthermore, the cytosolic domain of GPP130 interacts with the SNARE syntaxin 5 [27]. Syntaxin 5 is required for the early endosome-to-Golgi transport of STxB/STx1B (see below), and the GPP130-syntaxin 5 interaction is critical to support STxB/STx1B transport [27].…”

Section: Gpp130-the Endosomal Receptor For Stxb/stx1bmentioning

confidence: 99%

Targeting the Early Endosome-to-Golgi Transport of Shiga Toxins as a Therapeutic Strategy

Selyunin

Mukhopadhyay

2020

Toxins

View full text Add to dashboard Cite

Shiga toxin (STx) produced by Shigella and closely related Shiga toxin 1 and 2 (STx1 and STx2) synthesized by Shiga toxin-producing Escherichia coli (STEC) are bacterial AB5 toxins. All three toxins target kidney cells and may cause life-threatening renal disease. While Shigella infections can be treated with antibiotics, resistance is increasing. Moreover, antibiotic therapy is contraindicated for STEC, and there are no definitive treatments for STEC-induced disease. To exert cellular toxicity, STx, STx1, and STx2 must undergo retrograde trafficking to reach their cytosolic target, ribosomes. Direct transport from early endosomes to the Golgi apparatus is an essential step that allows the toxins to bypass degradative late endosomes and lysosomes. The essentiality of this transport step also makes it an ideal target for the development of small-molecule inhibitors of toxin trafficking as potential therapeutics. Here, we review the recent advances in understanding the molecular mechanisms of the early endosome-to-Golgi transport of STx, STx1, and STx2, as well as the development of small-molecule inhibitors of toxin trafficking that act at the endosome/Golgi interface.

show abstract

Section: Gpp130-the Endosomal Receptor For Stxb/stx1bmentioning

confidence: 99%

Targeting the Early Endosome-to-Golgi Transport of Shiga Toxins as a Therapeutic Strategy

Selyunin

Mukhopadhyay

2020

Toxins

View full text Add to dashboard Cite

show abstract

“…A small molecule inhibitor was tested to probe further the role of retrograde trafficking in HSV entry. Retro-2 blocks retrograde traffic from EE to the TGN, and consequently inhibits cytosolic uptake of Shiga and ricin toxins (58,(79)(80)(81)(82). Micromolar concentrations of retro-2 inhibits entry of human papillomavirus 16, BK virus, JC virus, and SV40, and transduction by adeno-associated virus serotype 2, all of which rely on retrograde transport mechanisms (58,73,75).…”

Section: Role Of Rab9 and Rab11 In Hsv-1 Entrymentioning

confidence: 99%

Herpes simplex virus entry by a non-conventional endocytic pathway

Tebaldi

Pritchard

Nicola

2020

Preprint

View full text Add to dashboard Cite

Herpes simplex virus 1 (HSV-1) causes significant morbidity and mortality in humans worldwide. HSV-1 enters epithelial cells via an endocytosis mechanism that is low pH-dependent. However, the precise intracellular pathway has not been identified, including the compartment where fusion occurs. In this study, we utilized a combination of molecular and pharmacological approaches to better characterize HSV entry by endocytosis. HSV-1 entry was unaltered in both cells treated with siRNA to Rab5 or Rab7 and cells expressing dominant-negative forms of these GTPases, suggesting entry is independent of the conventional endo-lysosomal network. The fungal metabolite brefeldin A (BFA) and the quinoline compound Golgicide A (GCA) inhibited HSV-1 entry via beta-galactosidase reporter assay and impaired incoming virus transport to the nuclear periphery, suggesting a role for trans Golgi network (TGN) functions and retrograde transport in HSV entry. Silencing of Rab9 or Rab11 GTPases, which are involved in the retrograde transport pathway, resulted in only a slight reduction in HSV infection. Together these results suggest that HSV enters host cells by an intracellular route independent of the lysosome-terminal endocytic pathway.IMPORTANCEHSV-1, the prototype alphaherpesvirus, is ubiquitous in the human population and causes lifelong infection that can be fatal in neonatal and immunocompromised individuals. HSV enters many cell types by endocytosis, including epithelial cells, the site of primary infection in the host. The intracellular itinerary for HSV entry remains unclear. We probed the potential involvement of several Rab GTPases in HSV-1 entry, and suggest that endocytic entry of HSV-1 is independent of the canonical lysosome-terminal pathway. A non-traditional endocytic route may be employed, such as one that intersects with the TGN. These results may lead to novel targets for intervention.

show abstract

“…We developed a pharmacological approach to block the maturation of L. infantum-containing phagosomes/PVs that occurs through heterotypic fusion with LEs and lysosomes. We used the small trafficking inhibitor Retro-2, known to block ricin and Shiga-toxin trafficking at the EEs-trans-Golgi network (TGN) interface [29,30]. For our time-dependent analysis, we took into account the previously [15][16][17][18] and above reported rapid insulation of L. infantum parasites within early tight-fitting phagosomes and subsequent mature tight-fitting PVs, also previously observed with tight-fitting PV-insulated L. major [27,31] and L. donovani [7,32].…”

Section: Impairment Of the Maturation Of Tight-fitting L Infantum Lementioning

confidence: 99%

“…Knowledge of the regulatory mechanisms that control such trafficking suggests that the inhibitory effects of Retro-2 are the result of its actions on various targets. Stx5-dependent effects [29,57,58] relate to a Retro-2-induced default of Stx5 localization to the TGN because it impairs Sec16A-dependent cycling of this SNAREs between the Golgi and the ER [30] and inhibits the ASNA1-mediated ER targeting and insertion of tail-anchored proteins [59]. Moreover, it promoted a default of the docking of EEs carrying Shiga toxin onto the TGN because the EE-associated Shiga toxin-trafficking chaperone GPP130 can no longer bind to its TGN-associated Stx5 receptor [30].…”

Section: Plos Neglected Tropical Diseasesmentioning

confidence: 99%

“…As a consequence, Retro-2 would be expected to have negative pleiotropic effects and yet, this is not the case [29]. Identical questions have been raised [64] concerning the blocking effect of Retro-2 at the ER [30,59]. For the effect on MT network it is possible that Retro-2 selectively affects one or several existing stable and dynamic sub-populations of αβ-tubulin dimers, or stable MTs with numerous post-translational modifications of tubulin, or various cellular pools of MTs [65,66].…”

Section: Plos Neglected Tropical Diseasesmentioning

confidence: 99%

See 1 more Smart Citation

The macrophage microtubule network acts as a key cellular controller of the intracellular fate of Leishmania infantum

2020

View full text Add to dashboard Cite

The parasitophorous vacuoles (PVs) that insulate Leishmania spp. in host macrophages are vacuolar compartments wherein promastigote forms differentiate into amastigote that are the replicative form of the parasite and are also more resistant to host responses. We revisited the biogenesis of tight-fitting PVs that insulate L. infantum in promastigote-infected macrophage-like RAW 264.7 cells by time-dependent confocal laser multidimensional imaging analysis. Pharmacological disassembly of the cellular microtubule network and silencing of the dynein gene led to an impaired interaction of L. infantum-containing phagosomes with late endosomes and lysosomes, resulting in the tight-fitting parasite-containing phagosomes never transforming into mature PVs. Analysis of the shape of the L. infantum parasite within PVs, showed that factors that impair promastigote-amastigote differentiation can also result in PVs whose maturation is arrested. These findings highlight the importance of the MT-dependent interaction of L. infantum-containing phagosomes with the host macrophage endolysosomal pathway to secure the intracellular fate of the parasite.

show abstract

Functional dissection of the retrograde Shiga toxin trafficking inhibitor Retro-2

Cited by 44 publications

References 57 publications

Targeting the Early Endosome-to-Golgi Transport of Shiga Toxins as a Therapeutic Strategy

Targeting the Early Endosome-to-Golgi Transport of Shiga Toxins as a Therapeutic Strategy

Herpes simplex virus entry by a non-conventional endocytic pathway

The macrophage microtubule network acts as a key cellular controller of the intracellular fate of Leishmania infantum

Contact Info

Product

Resources

About