{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:51:12Z","timestamp":1782280272006,"version":"3.54.5"},"posted":{"date-parts":[[2025,6,23]]},"group-title":"Microbiology","reference-count":45,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,6,23]],"date-time":"2025-06-23T00:00:00Z","timestamp":1750636800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,6,23]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The intracellular parasite\n                  <jats:italic>Toxoplasma gondii<\/jats:italic>\n                  enhances its dissemination to distant organs by hijacking infected leukocytes via a Trojan Horse mechanism. Upon infecting dendritic cells (DCs),\n                  <jats:italic>Toxoplasma<\/jats:italic>\n                  induces a hypermigratory phenotype characterized by podosome dissolution and formation of F-actin stress fibers. We previously showed that these cytoskeletal changes depend on the effector protein\n                  <jats:italic>Toxoplasma<\/jats:italic>\n                  WAVE complex-interacting protein (\n                  <jats:italic>Tg<\/jats:italic>\n                  WIP) secreted from parasites to infected leukocytes. Here, we identify the host adaptor proteins Non-catalytic region of tyrosine kinase adaptor protein 1 and 2 (Nck1\/2) and Growth factor receptor-bound protein 2 (Grb2) as direct\n                  <jats:italic>Tg<\/jats:italic>\n                  WIP interactors.\n                  <jats:italic>Tg<\/jats:italic>\n                  WIP mainly uses two distinct proline-rich regions (PRRs) to interact with Nck1 and Grb2. Mutating these PRRs abrogates\n                  <jats:italic>Tg<\/jats:italic>\n                  WIP binding to Nck1 and Grb2 and diminishes podosome dissolution and DC hypermotility. Furthermore, we show that\n                  <jats:italic>Tg<\/jats:italic>\n                  WIP directly interacts with the actin nucleation promoting factor WAVE Regulatory Complex (WRC) via a WRC-interacting receptor sequence (WIRS). Disrupting this interaction also influences actin cytoskeletal remodeling and DC hypermotility. Collectively, our data reveal that\n                  <jats:italic>Tg<\/jats:italic>\n                  WIP directly interacts with multiple actin regulators, including Nck1, Grb2, and the WRC, to remodel the actin cytoskeleton of the host cells, elucidating a key mechanism that\n                  <jats:italic>Toxoplasma<\/jats:italic>\n                  exploits to enhance host cell migration and dissemination.\n                <\/jats:p>","DOI":"10.1101\/2025.06.22.660983","type":"posted-content","created":{"date-parts":[[2025,6,23]],"date-time":"2025-06-23T23:55:16Z","timestamp":1750722916000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>Toxoplasma<\/i>\n                  effector\n                  <i>Tg<\/i>\n                  WIP hijacks dendritic cell actin and motility via Nck1\/Grb2 and the WAVE complex"],"prefix":"10.64898","author":[{"given":"Pavel","family":"Morales","sequence":"first","affiliation":[{"name":"Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel A.","family":"Kramer","sequence":"additional","affiliation":[{"name":"Roy J. Carver Department of Biochemistry, Biophysics & Molecular Biology, Iowa State University, Ames, IA, USA"},{"name":"Departments of Internal Medicine and Biophysics, University of Texas Southwestern Medical Center, Dallas, TX, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caroline","family":"de Moraes de Siqueira","sequence":"additional","affiliation":[{"name":"Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abbigale J.","family":"Brown","sequence":"additional","affiliation":[{"name":"Roy J. Carver Department of Biochemistry, Biophysics & Molecular Biology, Iowa State University, Ames, IA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lamba Omar","family":"Sangar\u00e9","sequence":"additional","affiliation":[{"name":"Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baoyu","family":"Chen","sequence":"additional","affiliation":[{"name":"Roy J. Carver Department of Biochemistry, Biophysics & Molecular Biology, Iowa State University, Ames, IA, USA"},{"name":"Departments of Internal Medicine and Biophysics, University of Texas Southwestern Medical Center, Dallas, TX, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0289-7109","authenticated-orcid":false,"given":"Jeroen P.J.","family":"Saeij","sequence":"additional","affiliation":[{"name":"Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026041209531704000_2025.06.22.660983v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/S0960-9822(02)00423-2"},{"key":"2026041209531704000_2025.06.22.660983v1.2","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.77220"},{"key":"2026041209531704000_2025.06.22.660983v1.3","doi-asserted-by":"publisher","DOI":"10.1002\/cam4.2640"},{"key":"2026041209531704000_2025.06.22.660983v1.4","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.8.4822-4826.2000"},{"key":"2026041209531704000_2025.06.22.660983v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2013.11.048"},{"key":"2026041209531704000_2025.06.22.660983v1.6","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-397924-7.00004-2"},{"key":"2026041209531704000_2025.06.22.660983v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2009.08.006"},{"issue":"September","key":"2026041209531704000_2025.06.22.660983v1.8","first-page":"107","article-title":"Hijacking Host Cell Highways: Manipulation of the Host Actin Cytoskeleton by Obligate Intracellular Bacterial Pathogens","volume":"6","year":"2016","journal-title":"Frontiers in Cellular and Infection Microbiology"},{"key":"2026041209531704000_2025.06.22.660983v1.9","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2005-02-0666"},{"key":"2026041209531704000_2025.06.22.660983v1.10","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-019-0504-8"},{"key":"2026041209531704000_2025.06.22.660983v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/nature00859"},{"key":"2026041209531704000_2025.06.22.660983v1.12","doi-asserted-by":"publisher","DOI":"10.1038\/nature08908"},{"key":"2026041209531704000_2025.06.22.660983v1.13","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7601092"},{"key":"2026041209531704000_2025.06.22.660983v1.14","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003051"},{"key":"2026041209531704000_2025.06.22.660983v1.15","doi-asserted-by":"publisher","DOI":"10.1038\/ncb0901-856"},{"key":"2026041209531704000_2025.06.22.660983v1.16","doi-asserted-by":"publisher","DOI":"10.1099\/00222615-30-4-302"},{"key":"2026041209531704000_2025.06.22.660983v1.17","first-page":"3.7.1","article-title":"Isolation of Dendritic Cells","volume":"3","year":"2009","journal-title":"Current Protocols in Immunology \/ Edited by John E. 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