Reconstruction of the Fig 4E panel I.
<div><p>The conoid is a dynamic, tubulin-based structure conserved across the Apicomplexa that undergoes extrusion during egress, gliding motility, and invasion in <i>Toxoplasma gondii</i>. This organelle traverses the apical polar ring (APR) in response to calcium waves and...
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2025
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| _version_ | 1849927642236583936 |
|---|---|
| author | Romuald Haase (22339777) |
| author2 | Bingjian Ren (22676577) Albert Tell i Puig (22676580) Alessandro Bonavoglia (19415718) Jean-Baptiste Marq (375239) Rémy Visentin (22676583) Nicolas Dos Santos Pacheco (22676586) Bohumil Maco (383443) Ricardo Mondragón-Flores (10737057) Oscar Vadas (2416228) Dominique Soldati-Favre (79026) |
| author2_role | author author author author author author author author author author |
| author_facet | Romuald Haase (22339777) Bingjian Ren (22676577) Albert Tell i Puig (22676580) Alessandro Bonavoglia (19415718) Jean-Baptiste Marq (375239) Rémy Visentin (22676583) Nicolas Dos Santos Pacheco (22676586) Bohumil Maco (383443) Ricardo Mondragón-Flores (10737057) Oscar Vadas (2416228) Dominique Soldati-Favre (79026) |
| author_role | author |
| dc.creator.none.fl_str_mv | Romuald Haase (22339777) Bingjian Ren (22676577) Albert Tell i Puig (22676580) Alessandro Bonavoglia (19415718) Jean-Baptiste Marq (375239) Rémy Visentin (22676583) Nicolas Dos Santos Pacheco (22676586) Bohumil Maco (383443) Ricardo Mondragón-Flores (10737057) Oscar Vadas (2416228) Dominique Soldati-Favre (79026) |
| dc.date.none.fl_str_mv | 2025-11-24T18:30:59Z |
| dc.identifier.none.fl_str_mv | 10.1371/journal.pbio.3003506.s010 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/media/Reconstruction_of_the_Fig_4E_panel_I_/30697341 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biophysics Biochemistry Microbiology Cell Biology Physiology Infectious Diseases Environmental Sciences not elsewhere classified Biological Sciences not elsewhere classified Physical Sciences not elsewhere classified toxoplasma gondii </ remains apically anchored heterologous expression systems div >< p apical polar ring biochemical analysis revealed proper conoid anchorage ensures conoid functionality less parasites revealed host cell invasion enable parasite motility controlling parasite motility although rng2 depletion intact conoid organelle organelle traverses intact protein conditional depletion biochemical properties detached conoid gliding motility unstable protein undergoes extrusion two structures strong candidate striking detachment still unclear rng2 tethers rng2 led rng2 emerges rhoptries follow resilient bridge protein localized plasma membrane pivotal protein likely facilitate large coiled immunoelectron microscopy functional activity enabling rng2 electron tomography dependent extrusion critical role concatenated assemblies comprehensive mutagenesis coil domains calcium waves |
| dc.title.none.fl_str_mv | Reconstruction of the Fig 4E panel I. |
| dc.type.none.fl_str_mv | Dataset Media info:eu-repo/semantics/publishedVersion dataset |
| description | <div><p>The conoid is a dynamic, tubulin-based structure conserved across the Apicomplexa that undergoes extrusion during egress, gliding motility, and invasion in <i>Toxoplasma gondii</i>. This organelle traverses the apical polar ring (APR) in response to calcium waves and plays a critical role in controlling parasite motility. While the actomyosin-dependent extrusion of the conoid is beginning to be elucidated, the mechanism by which it remains apically anchored to the APR is still unclear. RNG2, a protein localized to both the conoid and the APR, has emerged as a strong candidate for mediating this connection. Biochemical analysis revealed that RNG2 is an unstable protein, undergoing extensive proteolytic cleavage both in the parasite and in heterologous expression systems. Its biochemical properties, with the presence of large coiled-coil domains, likely facilitate the formation of concatenated assemblies, enabling RNG2 to serve as a dynamic and resilient bridge between the conoid and the APR. Using a combination of iterative ultrastructure expansion microscopy and immunoelectron microscopy, we confirmed the localization of RNG2 to the 22 tethering elements bridging the APR and the conoid. Conditional depletion of RNG2 led to the striking detachment of the intact conoid organelle from the APR, supporting an essential role for RNG2 as a tether. Cryo-electron tomography of conoid-less parasites revealed that, in the absence of RNG2, the apical vesicle remains anchored to the plasma membrane, while the rhoptries follow the detached conoid. Although RNG2 depletion only mildly reduces microneme secretion, the parasites are immotile and exhibit impaired rhoptry discharge, highlighting the critical role of proper conoid anchorage in motility and host cell invasion. Comprehensive mutagenesis of RNG2 identified distinct regions responsible for binding to the conoid and the APR, and demonstrated that the full-length, intact protein is essential for bridging these two structures and for its functional activity. Altogether, RNG2 emerges as a pivotal protein that ensures conoid functionality and coordination in Coccidia.</p></div> |
| eu_rights_str_mv | openAccess |
| id | Manara_e2e009cc76a9fdeacb2254bd25e50674 |
| identifier_str_mv | 10.1371/journal.pbio.3003506.s010 |
| network_acronym_str | Manara |
| network_name_str | ManaraRepo |
| oai_identifier_str | oai:figshare.com:article/30697341 |
| publishDate | 2025 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Reconstruction of the Fig 4E panel I.Romuald Haase (22339777)Bingjian Ren (22676577)Albert Tell i Puig (22676580)Alessandro Bonavoglia (19415718)Jean-Baptiste Marq (375239)Rémy Visentin (22676583)Nicolas Dos Santos Pacheco (22676586)Bohumil Maco (383443)Ricardo Mondragón-Flores (10737057)Oscar Vadas (2416228)Dominique Soldati-Favre (79026)BiophysicsBiochemistryMicrobiologyCell BiologyPhysiologyInfectious DiseasesEnvironmental Sciences not elsewhere classifiedBiological Sciences not elsewhere classifiedPhysical Sciences not elsewhere classifiedtoxoplasma gondii </remains apically anchoredheterologous expression systemsdiv >< papical polar ringbiochemical analysis revealedproper conoid anchorageensures conoid functionalityless parasites revealedhost cell invasionenable parasite motilitycontrolling parasite motilityalthough rng2 depletionintact conoid organelleorganelle traversesintact proteinconditional depletionbiochemical propertiesdetached conoidgliding motilityunstable proteinundergoes extrusiontwo structuresstrong candidatestriking detachmentstill unclearrng2 tethersrng2 ledrng2 emergesrhoptries followresilient bridgeprotein localizedplasma membranepivotal proteinlikely facilitatelarge coiledimmunoelectron microscopyfunctional activityenabling rng2electron tomographydependent extrusioncritical roleconcatenated assembliescomprehensive mutagenesiscoil domainscalcium waves<div><p>The conoid is a dynamic, tubulin-based structure conserved across the Apicomplexa that undergoes extrusion during egress, gliding motility, and invasion in <i>Toxoplasma gondii</i>. This organelle traverses the apical polar ring (APR) in response to calcium waves and plays a critical role in controlling parasite motility. While the actomyosin-dependent extrusion of the conoid is beginning to be elucidated, the mechanism by which it remains apically anchored to the APR is still unclear. RNG2, a protein localized to both the conoid and the APR, has emerged as a strong candidate for mediating this connection. Biochemical analysis revealed that RNG2 is an unstable protein, undergoing extensive proteolytic cleavage both in the parasite and in heterologous expression systems. Its biochemical properties, with the presence of large coiled-coil domains, likely facilitate the formation of concatenated assemblies, enabling RNG2 to serve as a dynamic and resilient bridge between the conoid and the APR. Using a combination of iterative ultrastructure expansion microscopy and immunoelectron microscopy, we confirmed the localization of RNG2 to the 22 tethering elements bridging the APR and the conoid. Conditional depletion of RNG2 led to the striking detachment of the intact conoid organelle from the APR, supporting an essential role for RNG2 as a tether. Cryo-electron tomography of conoid-less parasites revealed that, in the absence of RNG2, the apical vesicle remains anchored to the plasma membrane, while the rhoptries follow the detached conoid. Although RNG2 depletion only mildly reduces microneme secretion, the parasites are immotile and exhibit impaired rhoptry discharge, highlighting the critical role of proper conoid anchorage in motility and host cell invasion. Comprehensive mutagenesis of RNG2 identified distinct regions responsible for binding to the conoid and the APR, and demonstrated that the full-length, intact protein is essential for bridging these two structures and for its functional activity. Altogether, RNG2 emerges as a pivotal protein that ensures conoid functionality and coordination in Coccidia.</p></div>2025-11-24T18:30:59ZDatasetMediainfo:eu-repo/semantics/publishedVersiondataset10.1371/journal.pbio.3003506.s010https://figshare.com/articles/media/Reconstruction_of_the_Fig_4E_panel_I_/30697341CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/306973412025-11-24T18:30:59Z |
| spellingShingle | Reconstruction of the Fig 4E panel I. Romuald Haase (22339777) Biophysics Biochemistry Microbiology Cell Biology Physiology Infectious Diseases Environmental Sciences not elsewhere classified Biological Sciences not elsewhere classified Physical Sciences not elsewhere classified toxoplasma gondii </ remains apically anchored heterologous expression systems div >< p apical polar ring biochemical analysis revealed proper conoid anchorage ensures conoid functionality less parasites revealed host cell invasion enable parasite motility controlling parasite motility although rng2 depletion intact conoid organelle organelle traverses intact protein conditional depletion biochemical properties detached conoid gliding motility unstable protein undergoes extrusion two structures strong candidate striking detachment still unclear rng2 tethers rng2 led rng2 emerges rhoptries follow resilient bridge protein localized plasma membrane pivotal protein likely facilitate large coiled immunoelectron microscopy functional activity enabling rng2 electron tomography dependent extrusion critical role concatenated assemblies comprehensive mutagenesis coil domains calcium waves |
| status_str | publishedVersion |
| title | Reconstruction of the Fig 4E panel I. |
| title_full | Reconstruction of the Fig 4E panel I. |
| title_fullStr | Reconstruction of the Fig 4E panel I. |
| title_full_unstemmed | Reconstruction of the Fig 4E panel I. |
| title_short | Reconstruction of the Fig 4E panel I. |
| title_sort | Reconstruction of the Fig 4E panel I. |
| topic | Biophysics Biochemistry Microbiology Cell Biology Physiology Infectious Diseases Environmental Sciences not elsewhere classified Biological Sciences not elsewhere classified Physical Sciences not elsewhere classified toxoplasma gondii </ remains apically anchored heterologous expression systems div >< p apical polar ring biochemical analysis revealed proper conoid anchorage ensures conoid functionality less parasites revealed host cell invasion enable parasite motility controlling parasite motility although rng2 depletion intact conoid organelle organelle traverses intact protein conditional depletion biochemical properties detached conoid gliding motility unstable protein undergoes extrusion two structures strong candidate striking detachment still unclear rng2 tethers rng2 led rng2 emerges rhoptries follow resilient bridge protein localized plasma membrane pivotal protein likely facilitate large coiled immunoelectron microscopy functional activity enabling rng2 electron tomography dependent extrusion critical role concatenated assemblies comprehensive mutagenesis coil domains calcium waves |