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This process requires the prior recognition and binding of the host cell by proteins released from another set of parasite organelles, the micronemes. However, the host-parasite interactions required for rhoptry discharge are largely unknown. Here we performed a host-cell directed genome-wide screen for host factors required for rhoptry discharge from\n                  <jats:italic>Toxoplasma gondii,<\/jats:italic>\n                  the causative agent of toxoplasmosis. The screen identified host N-glycosylation and cholesterol biosynthesis as pathways required for normal rhoptry discharge. A trimeric microneme complex, MIC1\/4\/6, interfaces with both pathways by binding host N-glycans to cluster proteins in a process dependent on host plasma membrane cholesterol. The process can be inhibited by depletion of host cholesterol or competition with exogenous glycans. This clustering of host factors by MIC1\/4\/6 likely prepares the host membrane for rhoptry discharge, delineating a new step in the\n                  <jats:italic>Toxoplasma<\/jats:italic>\n                  invasion process.\n                <\/jats:p>","DOI":"10.1101\/2025.10.16.682961","type":"posted-content","created":{"date-parts":[[2025,10,17]],"date-time":"2025-10-17T20:05:14Z","timestamp":1760731514000},"source":"Crossref","is-referenced-by-count":1,"title":["Clustering of host N-glycans licenses\n                  <i>Toxoplasma<\/i>\n                  rhoptry discharge"],"prefix":"10.64898","author":[{"given":"Dylan","family":"Valleau","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mudita","family":"Goyal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Justin M.","family":"Roberts","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frances","family":"Male","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gary","family":"Ward","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5237-1095","authenticated-orcid":false,"given":"Sebastian","family":"Lourido","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025102400052634000_2025.10.16.682961v2.1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1004670"},{"key":"2025102400052634000_2025.10.16.682961v2.2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.9.5379-5387.2005"},{"key":"2025102400052634000_2025.10.16.682961v2.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2006.06.017"},{"key":"2025102400052634000_2025.10.16.682961v2.4","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1186\/s12915-016-0316-8","article-title":"Genetic impairment of parasite myosin motors uncovers the contribution of host cell membrane dynamics to Toxoplasma invasion forces","volume":"14","year":"2016","journal-title":"BMC Biol"},{"key":"2025102400052634000_2025.10.16.682961v2.5","first-page":"114","article-title":"Sequential protein secretion from three distinct organelles of Toxoplasma gondii accompanies invasion of human fibroblasts","volume":"73","year":"1997","journal-title":"Eur. 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