{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T07:58:14Z","timestamp":1790668694731,"version":"4.1.0"},"reference-count":32,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,12,20]],"date-time":"2017-12-20T00:00:00Z","timestamp":1513728000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,12,20]],"date-time":"2017-12-20T00:00:00Z","timestamp":1513728000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Apicomplexa parasites, including <jats:italic>Toxoplasma<\/jats:italic> and <jats:italic>Plasmodium<\/jats:italic> species, possess a unique invasion mechanism that involves a tight apposition between the parasite and the host plasma membranes, called \u201cmoving junction\u201d (MJ). The MJ is formed by the assembly of the microneme protein AMA1, exposed at the surface of the parasite, and the parasite rhoptry neck (RON) protein RON2, exposed at the surface of the host cell. In the host cell, RON2 is associated with three additional parasite RON proteins, RON4, RON5 and RON8. Here we describe RON4<jats:sub>L1<\/jats:sub>, an additional member of the MJ complex in <jats:italic>Toxoplasma<\/jats:italic>. RON4<jats:sub>L1<\/jats:sub> displays some sequence similarity with RON4 and is cleaved at the C-terminal end before reaching the rhoptry neck. Upon secretion during invasion, RON4<jats:sub>L1<\/jats:sub> is associated with MJ and targeted to the cytosolic face of the host membrane. We generated a <jats:italic>RON4<\/jats:italic><jats:sub>\n                  <jats:italic>L1<\/jats:italic>\n                <\/jats:sub> knock-out cell line and showed that it is not essential for the lytic cycle <jats:italic>in vi<\/jats:italic>tro, although mutant parasites kill mice less efficiently. Similarly to RON8, RON4<jats:sub>L1<\/jats:sub> is a coccidian-specific protein and its traffic to the MJ is not affected in absence of RON2, RON4 and RON5, suggesting the co-existence of independent MJ complexes in tachyzoite of <jats:italic>Toxoplasma<\/jats:italic>.<\/jats:p>","DOI":"10.1038\/s41598-017-18010-9","type":"journal-article","created":{"date-parts":[[2017,12,14]],"date-time":"2017-12-14T15:26:16Z","timestamp":1513265176000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["RON4L1 is a new member of the moving junction complex in Toxoplasma gondii"],"prefix":"10.1038","volume":"7","author":[{"given":"Amandine","family":"Gu\u00e9rin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiba","family":"El Hajj","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diana","family":"Penarete-Vargas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S\u00e9bastien","family":"Besteiro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0962-0156","authenticated-orcid":false,"given":"Maryse","family":"Lebrun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,12,20]]},"reference":[{"key":"18010_CR1","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1083\/jcb.77.1.72","volume":"77","author":"M Aikawa","year":"1978","unstructured":"Aikawa, M., Miller, L. H., Johnson, J. & Rabbege, J. Erythrocyte entry by malarial parasites. A moving junction between erythrocyte and parasite. J Cell Biol \n                           77, 72\u201382 (1978).","journal-title":"J Cell Biol"},{"key":"18010_CR2","doi-asserted-by":"publisher","first-page":"e17","DOI":"10.1371\/journal.ppat.0010017","volume":"1","author":"DL Alexander","year":"2005","unstructured":"Alexander, D. L., Mital, J., Ward, G. E., Bradley, P. & Boothroyd, J. C. Identification of the Moving Junction Complex of Toxoplasma gondii: A Collaboration between Distinct Secretory Organelles. PLoS Pathog \n                           1, e17 (2005).","journal-title":"PLoS Pathog"},{"key":"18010_CR3","doi-asserted-by":"publisher","first-page":"1823","DOI":"10.1111\/j.1462-5822.2005.00646.x","volume":"7","author":"M Lebrun","year":"2005","unstructured":"Lebrun, M. et al. The rhoptry neck protein RON4 relocalizes at the moving junction during Toxoplasma gondii invasion. Cell Microbiol \n                           7, 1823\u20131833 (2005).","journal-title":"Cell Microbiol"},{"key":"18010_CR4","doi-asserted-by":"publisher","first-page":"e1000309","DOI":"10.1371\/journal.ppat.1000309","volume":"5","author":"S Besteiro","year":"2009","unstructured":"Besteiro, S., Michelin, A., Poncet, J., Dubremetz, J. F. & Lebrun, M. Export of a Toxoplasma gondii rhoptry neck protein complex at the host cell membrane to form the moving junction during invasion. PLoS Pathog \n                           5, e1000309 (2009).","journal-title":"PLoS Pathog"},{"key":"18010_CR5","doi-asserted-by":"publisher","first-page":"590","DOI":"10.1111\/j.1462-5822.2008.01276.x","volume":"11","author":"KW Straub","year":"2009","unstructured":"Straub, K. W., Cheng, S. J., Sohn, C. S. & Bradley, P. J. Novel components of the Apicomplexan moving junction reveal conserved and coccidia-restricted elements. Cell Microbiol \n                           11, 590\u2013603 (2009).","journal-title":"Cell Microbiol"},{"key":"18010_CR6","doi-asserted-by":"publisher","first-page":"841","DOI":"10.1111\/j.1462-5822.2007.00909.x","volume":"9","author":"JF Dubremetz","year":"2007","unstructured":"Dubremetz, J. F. Rhoptries are major players in Toxoplasma gondii invasion and host cell interaction. Cell Microbiol \n                           9, 841\u2013848 (2007).","journal-title":"Cell Microbiol"},{"key":"18010_CR7","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1016\/0166-6851(94)90096-5","volume":"67","author":"DL Narum","year":"1994","unstructured":"Narum, D. L. & Thomas, A. W. Differential localization of full-length and processed forms of PF83\/AMA-1 an apical membrane antigen of Plasmodium falciparum merozoites. Mol Biochem Parasitol \n                           67, 59\u201368 (1994).","journal-title":"Mol Biochem Parasitol"},{"key":"18010_CR8","doi-asserted-by":"publisher","first-page":"e1001276","DOI":"10.1371\/journal.ppat.1001276","volume":"7","author":"M Lamarque","year":"2011","unstructured":"Lamarque, M. et al. The RON2-AMA1 interaction is a critical step in moving junction-dependent invasion by apicomplexan parasites. PLoS Pathog \n                           7, e1001276 (2011).","journal-title":"PLoS Pathog"},{"key":"18010_CR9","doi-asserted-by":"publisher","first-page":"e1001282","DOI":"10.1371\/journal.ppat.1001282","volume":"7","author":"JS Tyler","year":"2011","unstructured":"Tyler, J. S. & Boothroyd, J. C. The C-terminus of Toxoplasma RON2 provides the crucial link between AMA1 and the host-associated invasion complex. PLoS Pathog \n                           7, e1001282, https:\/\/doi.org\/10.1371\/journal.ppat.1001282 (2011).","journal-title":"PLoS Pathog"},{"key":"18010_CR10","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1126\/science.1204988","volume":"333","author":"ML Tonkin","year":"2011","unstructured":"Tonkin, M. L. et al. Host cell invasion by apicomplexan parasites: insights from the co-structure of AMA1 with a RON2 peptide. Science \n                           333, 463\u2013467 (2011).","journal-title":"Science"},{"key":"18010_CR11","doi-asserted-by":"publisher","first-page":"e1002755","DOI":"10.1371\/journal.ppat.1002755","volume":"8","author":"B Vulliez-Le Normand","year":"2012","unstructured":"Vulliez-Le Normand, B. et al. Structural and functional insights into the malaria parasite moving junction complex. PLoS pathogens \n                           8, e1002755 (2012).","journal-title":"PLoS pathogens"},{"key":"18010_CR12","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms5098","volume":"5","author":"MH Lamarque","year":"2014","unstructured":"Lamarque, M. H. et al. Plasticity and redundancy among AMA-RON pairs ensure host cell entry of Toxoplasma parasites. Nat Commun \n                           5, 4098, https:\/\/doi.org\/10.1038\/ncomms5098 (2014).","journal-title":"Nat Commun"},{"key":"18010_CR13","doi-asserted-by":"publisher","first-page":"1358","DOI":"10.1038\/s41564-017-0018-1","volume":"2","author":"A Guerin","year":"2017","unstructured":"Guerin, A. et al. Efficient invasion by Toxoplasma depends on the subversion of host protein networks. Nat Microbiol \n                           2, 1358\u20131366, https:\/\/doi.org\/10.1038\/s41564-017-0018-1 (2017).","journal-title":"Nat Microbiol"},{"key":"18010_CR14","doi-asserted-by":"publisher","unstructured":"Wang, M. et al. The moving junction protein RON4, although not critical, facilitates host cell invasion and stabilizes MJ members. Parasitology, 1\u20138, https:\/\/doi.org\/10.1017\/S0031182017000968 (2017).","DOI":"10.1017\/S0031182017000968"},{"key":"18010_CR15","doi-asserted-by":"publisher","first-page":"e1002007","DOI":"10.1371\/journal.ppat.1002007","volume":"7","author":"KW Straub","year":"2011","unstructured":"Straub, K. W., Peng, E. D., Hajagos, B. E., Tyler, J. S. & Bradley, P. J. The moving junction protein RON8 facilitates firm attachment and host cell invasion in Toxoplasma gondii. PLoS Pathog \n                           7, e1002007, https:\/\/doi.org\/10.1371\/journal.ppat.1002007 (2011).","journal-title":"PLoS Pathog"},{"key":"18010_CR16","doi-asserted-by":"publisher","first-page":"e1004025","DOI":"10.1371\/journal.ppat.1004025","volume":"10","author":"JR Beck","year":"2014","unstructured":"Beck, J. R., Chen, A. L., Kim, E. W. & Bradley, P. J. RON5 is critical for organization and function of the Toxoplasma moving junction complex. PLoS Pathog \n                           10, e1004025, https:\/\/doi.org\/10.1371\/journal.ppat.1004025 (2014).","journal-title":"PLoS Pathog"},{"key":"18010_CR17","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms3552","volume":"4","author":"DY Bargieri","year":"2013","unstructured":"Bargieri, D. Y. et al. Apical membrane antigen 1 mediates apicomplexan parasite attachment but is dispensable for host cell invasion. Nat Commun \n                           4, 2552, https:\/\/doi.org\/10.1038\/ncomms3552 (2013).","journal-title":"Nat Commun"},{"key":"18010_CR18","doi-asserted-by":"publisher","first-page":"591","DOI":"10.1016\/j.chom.2011.10.012","volume":"10","author":"D Giovannini","year":"2011","unstructured":"Giovannini, D. et al. Independent roles of apical membrane antigen 1 and rhoptry neck proteins during host cell invasion by apicomplexa. Cell Host Microbe \n                           10, 591\u2013602, https:\/\/doi.org\/10.1016\/j.chom.2011.10.012 (2011).","journal-title":"Cell Host Microbe"},{"key":"18010_CR19","doi-asserted-by":"publisher","first-page":"398","DOI":"10.1073\/pnas.1515898113","volume":"113","author":"ML Parker","year":"2016","unstructured":"Parker, M. L. et al. Dissecting the interface between apicomplexan parasite and host cell: Insights from a divergent AMA-RON2 pair. Proc Natl Acad Sci USA \n                           113, 398\u2013403, https:\/\/doi.org\/10.1073\/pnas.1515898113 (2016).","journal-title":"Proc Natl Acad Sci USA"},{"key":"18010_CR20","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1038\/nrmicro1800","volume":"6","author":"JC Boothroyd","year":"2008","unstructured":"Boothroyd, J. C. & Dubremetz, J. F. Kiss and spit: the dual roles of Toxoplasma rhoptries. Nat Rev Microbiol \n                           6, 79\u201388 (2008).","journal-title":"Nat Rev Microbiol"},{"key":"18010_CR21","doi-asserted-by":"publisher","first-page":"e12354","DOI":"10.1371\/journal.pone.0012354","volume":"5","author":"MS Behnke","year":"2010","unstructured":"Behnke, M. S. et al. Coordinated progression through two subtranscriptomes underlies the tachyzoite cycle of Toxoplasma gondii. PLoS One \n                           5, e12354, https:\/\/doi.org\/10.1371\/journal.pone.0012354 (2010).","journal-title":"PLoS One"},{"key":"18010_CR22","doi-asserted-by":"publisher","first-page":"e29998","DOI":"10.1371\/journal.pone.0029998","volume":"7","author":"HM Fritz","year":"2012","unstructured":"Fritz, H. M. et al. Transcriptomic analysis of toxoplasma development reveals many novel functions and structures specific to sporozoites and oocysts. PLoS One \n                           7, e29998, https:\/\/doi.org\/10.1371\/journal.pone.0029998 (2012).","journal-title":"PLoS One"},{"key":"18010_CR23","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2164-15-350","volume":"15","author":"MS Behnke","year":"2014","unstructured":"Behnke, M. S., Zhang, T. P., Dubey, J. P. & Sibley, L. D. Toxoplasma gondii merozoite gene expression analysis with comparison to the life cycle discloses a unique expression state during enteric development. BMC Genomics \n                           15, 350, https:\/\/doi.org\/10.1186\/1471-2164-15-350 (2014).","journal-title":"BMC Genomics"},{"key":"18010_CR24","doi-asserted-by":"publisher","first-page":"530","DOI":"10.1128\/EC.00358-08","volume":"8","author":"MH Huynh","year":"2009","unstructured":"Huynh, M. H. & Carruthers, V. B. Tagging of endogenous genes in a Toxoplasma gondii strain lacking Ku80. Eukaryot Cell \n                           8, 530\u2013539 (2009).","journal-title":"Eukaryot Cell"},{"key":"18010_CR25","doi-asserted-by":"publisher","first-page":"1320","DOI":"10.1128\/EC.3.5.1320-1330.2004","volume":"3","author":"KL Carey","year":"2004","unstructured":"Carey, K. L., Jongco, A. M., Kim, K. & Ward, G. E. The Toxoplasma gondii rhoptry protein ROP4 is secreted into the parasitophorous vacuole and becomes phosphorylated in infected cells. Eukaryot Cell \n                           3, 1320\u20131330 (2004).","journal-title":"Eukaryot Cell"},{"key":"18010_CR26","first-page":"114","volume":"73","author":"VB Carruthers","year":"1997","unstructured":"Carruthers, V. B. & Sibley, L. D. Sequential protein secretion from three distinct organelles of Toxoplasma gondii accompanies invasion of human fibroblasts. Eur J Cell Biol \n                           73, 114\u2013123 (1997).","journal-title":"Eur J Cell Biol"},{"key":"18010_CR27","doi-asserted-by":"publisher","first-page":"e70637","DOI":"10.1371\/journal.pone.0070637","volume":"8","author":"A Poukchanski","year":"2013","unstructured":"Poukchanski, A. et al. Toxoplasma gondii Sporozoites Invade Host Cells Using Two Novel Paralogues of RON2 and AMA1. PLoS One \n                           8, e70637 (2013).","journal-title":"PLoS One"},{"key":"18010_CR28","doi-asserted-by":"crossref","unstructured":"Frenal, K., Dubremetz, J. F., Lebrun, M. & soldati-Favre, D. Gliding motility powers invasion and egress in Apicomplexa. Nature Reviews in press (2017).","DOI":"10.1038\/nrmicro.2017.86"},{"key":"18010_CR29","doi-asserted-by":"publisher","first-page":"1019","DOI":"10.1128\/EC.00413-07","volume":"7","author":"H El Hajj","year":"2008","unstructured":"El Hajj, H. et al. Molecular signals in the trafficking of Toxoplasma gondii protein MIC3 to the micronemes. Eukaryot Cell \n                           7, 1019\u20131028 (2008).","journal-title":"Eukaryot Cell"},{"key":"18010_CR30","doi-asserted-by":"publisher","first-page":"911","DOI":"10.1126\/science.8235614","volume":"262","author":"K Kim","year":"1993","unstructured":"Kim, K., Soldati, D. & Boothroyd, J. C. Gene replacement in Toxoplasma gondii with chloramphenicol acetyltransferase as selectable marker. Science \n                           262, 911\u2013914 (1993).","journal-title":"Science"},{"key":"18010_CR31","doi-asserted-by":"publisher","first-page":"559","DOI":"10.1111\/cmi.12383","volume":"17","author":"W Daher","year":"2015","unstructured":"Daher, W. et al. Lipid kinases are essential for apicoplast homeostasis in Toxoplasma gondii. Cell Microbiol \n                           17, 559\u2013578, https:\/\/doi.org\/10.1111\/cmi.12383 (2015).","journal-title":"Cell Microbiol"},{"issue":"Pt 5","key":"18010_CR32","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1242\/jcs.88.5.669","volume":"88","author":"PL McNeil","year":"1987","unstructured":"McNeil, P. L. & Warder, E. Glass beads load macromolecules into living cells. J Cell Sci \n                           88(Pt 5), 669\u2013678 (1987).","journal-title":"J Cell Sci"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-18010-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-18010-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-18010-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T17:07:21Z","timestamp":1671815241000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-18010-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,12,20]]},"references-count":32,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["18010"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-18010-9","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,12,20]]},"assertion":[{"value":"17 August 2017","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"5 December 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"20 December 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"17907"}}