RNG2 is functional only as intact, full-length protein.
<p><b>(A)</b> Representation of the RNG2 3 processed products originally identified in insect cells (155kDa, 90kDa, 60kDa) expressed in the Δ<i>rng2</i> background. <b>(B)</b> Western blot analysis of the expression of RNG2 variants expressed in the Δ<i&g...
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2025
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| _version_ | 1849927642174717952 |
|---|---|
| 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 | 2025-11-24T18:31:14Z |
| dc.identifier.none.fl_str_mv | 10.1371/journal.pbio.3003506.g007 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/figure/RNG2_is_functional_only_as_intact_full-length_protein_/30697377 |
| 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 | RNG2 is functional only as intact, full-length protein. |
| dc.type.none.fl_str_mv | Image Figure info:eu-repo/semantics/publishedVersion image |
| description | <p><b>(A)</b> Representation of the RNG2 3 processed products originally identified in insect cells (155kDa, 90kDa, 60kDa) expressed in the Δ<i>rng2</i> background. <b>(B)</b> Western blot analysis of the expression of RNG2 variants expressed in the Δ<i>rng2</i> background. <b>(C)</b> U-ExM images of the localization of the RNG2 variants. Scale bar = 3 μm. <b>(D)</b> Quantification of the plaque sizes created by the different RNG2 strains via a plaque assay. Ten plaques were measured per biological replicate. Three biological replicates were evaluated. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s008" target="_blank">S2 Data</a>. <b>(E)</b> Quantification of the percentage of invasion by the different RNG2 strains in the invasion assay. One hundred parasites were measured per biological replicate. Three biological replicates were evaluated. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s008" target="_blank">S2 Data</a>. <b>(F)</b> Quantification of the percentages of conoid retraction, extrusion, and detachment by the different RNG2 strains in the conoid detachment assay. One hundred parasites were measured per biological replicate. Three biological replicates were evaluated. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s008" target="_blank">S2 Data</a>. <b>(G)</b> Design of the experiment to test if different fragments of RNG2 can cooperate to form a functional tether. <b>(H)</b> Conoid detachment assessed by U-ExM in transiently transfected parasites treated for 48h under auxin to down-regulate the endogenous RNG2-mAID. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s007" target="_blank">S1 Data</a>. <b>(I)</b> Representation of the skip peptide (P2A) variants expressed in the RNG2-mAID-HA strain. <b>(J)</b> Western blot analysis of the expression and skipping pattern verifying the successful skipping of ribosome on the P2A sequence. <b>(K)</b> U-ExM images on auxin treated parasite to deplete the endogenous RNG2, showing the localization of both P2A variants and their ability to maintain the conoid at the apical pole. <b>(L)</b> Quantification of the plaque sizes created by RNG2 P2A variants via a plaque assay. Ten plaques were measured per biological replicate. Three biological replicates were evaluated. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s008" target="_blank">S2 Data</a>.</p> |
| eu_rights_str_mv | openAccess |
| id | Manara_a9e7816bd066901e013b1b60cb9efa13 |
| identifier_str_mv | 10.1371/journal.pbio.3003506.g007 |
| network_acronym_str | Manara |
| network_name_str | ManaraRepo |
| oai_identifier_str | oai:figshare.com:article/30697377 |
| publishDate | 2025 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | RNG2 is functional only as intact, full-length protein.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<p><b>(A)</b> Representation of the RNG2 3 processed products originally identified in insect cells (155kDa, 90kDa, 60kDa) expressed in the Δ<i>rng2</i> background. <b>(B)</b> Western blot analysis of the expression of RNG2 variants expressed in the Δ<i>rng2</i> background. <b>(C)</b> U-ExM images of the localization of the RNG2 variants. Scale bar = 3 μm. <b>(D)</b> Quantification of the plaque sizes created by the different RNG2 strains via a plaque assay. Ten plaques were measured per biological replicate. Three biological replicates were evaluated. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s008" target="_blank">S2 Data</a>. <b>(E)</b> Quantification of the percentage of invasion by the different RNG2 strains in the invasion assay. One hundred parasites were measured per biological replicate. Three biological replicates were evaluated. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s008" target="_blank">S2 Data</a>. <b>(F)</b> Quantification of the percentages of conoid retraction, extrusion, and detachment by the different RNG2 strains in the conoid detachment assay. One hundred parasites were measured per biological replicate. Three biological replicates were evaluated. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s008" target="_blank">S2 Data</a>. <b>(G)</b> Design of the experiment to test if different fragments of RNG2 can cooperate to form a functional tether. <b>(H)</b> Conoid detachment assessed by U-ExM in transiently transfected parasites treated for 48h under auxin to down-regulate the endogenous RNG2-mAID. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s007" target="_blank">S1 Data</a>. <b>(I)</b> Representation of the skip peptide (P2A) variants expressed in the RNG2-mAID-HA strain. <b>(J)</b> Western blot analysis of the expression and skipping pattern verifying the successful skipping of ribosome on the P2A sequence. <b>(K)</b> U-ExM images on auxin treated parasite to deplete the endogenous RNG2, showing the localization of both P2A variants and their ability to maintain the conoid at the apical pole. <b>(L)</b> Quantification of the plaque sizes created by RNG2 P2A variants via a plaque assay. Ten plaques were measured per biological replicate. Three biological replicates were evaluated. The data underlying this figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003506#pbio.3003506.s008" target="_blank">S2 Data</a>.</p>2025-11-24T18:31:14ZImageFigureinfo:eu-repo/semantics/publishedVersionimage10.1371/journal.pbio.3003506.g007https://figshare.com/articles/figure/RNG2_is_functional_only_as_intact_full-length_protein_/30697377CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/306973772025-11-24T18:31:14Z |
| spellingShingle | RNG2 is functional only as intact, full-length protein. 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 | RNG2 is functional only as intact, full-length protein. |
| title_full | RNG2 is functional only as intact, full-length protein. |
| title_fullStr | RNG2 is functional only as intact, full-length protein. |
| title_full_unstemmed | RNG2 is functional only as intact, full-length protein. |
| title_short | RNG2 is functional only as intact, full-length protein. |
| title_sort | RNG2 is functional only as intact, full-length protein. |
| 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 |