{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T12:13:59Z","timestamp":1785413639199,"version":"3.56.0"},"posted":{"date-parts":[[2025,4,23]]},"group-title":"Microbiology","reference-count":59,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,4,23]],"date-time":"2025-04-23T00:00:00Z","timestamp":1745366400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"award":["AI069314"],"award-info":[{"award-number":["AI069314"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"award":["AI103638"],"award-info":[{"award-number":["AI103638"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01 AI076276"],"award-info":[{"award-number":["R01 AI076276"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01 AI125565"],"award-info":[{"award-number":["R01 AI125565"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Intramural research funds, Uniformed Services University of the Health Sciences, USA","award":["C073384316"],"award-info":[{"award-number":["C073384316"]}]},{"name":"Intramural research fund, Center for Quantitative Biology, Princeton University","award":["P50 GM071508"],"award-info":[{"award-number":["P50 GM071508"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,4,23]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Sexual commitment in\n                  <jats:italic>Plasmodium<\/jats:italic>\n                  parasites is essential for malaria transmission, yet much remains unknown about the underlying signaling events initiating sexual conversion in a subpopulation of parasites. We discovered a single valine (V\n                  <jats:sub>2163<\/jats:sub>\n                  ) to leucine (L\n                  <jats:sub>2163<\/jats:sub>\n                  ) mutation in an Apetala 2 (AP2) transcription factor required for\n                  <jats:italic>P. falciparum<\/jats:italic>\n                  gametocytogenesis,\n                  <jats:italic>ap2-g<\/jats:italic>\n                  that abrogates sexual differentiation and confirmed this with forward and reverse mutation editing. Mutated AP2-G.L\n                  <jats:sub>2163<\/jats:sub>\n                  does not bind the\n                  <jats:italic>ap2-g<\/jats:italic>\n                  consensus motif, GnGTAC, or stimulate AP2-G-dependent gene transcription including autoregulation. We then used AP2-G.L\n                  <jats:sub>2163<\/jats:sub>\n                  parasite lines as tools to demonstrate the critical role of GDV1 in the initial activation of the silent\n                  <jats:italic>ap2-g<\/jats:italic>\n                  locus during the trophozoite to schizont transition in the absence of functional AP2-G and its autoregulation. Additionally, we show that AP2-G.V is required for MSRP1 expression, which can be used to distinguish early and late sexually committed schizonts. Together this work demonstrates that valine\n                  <jats:sub>2163<\/jats:sub>\n                  in AP2-G plays a critical role in DNA binding, highlighting the functional importance of this specific region for malaria transmission as well as the critical role of GDV1 in the initial activation of\n                  <jats:italic>ap2-g<\/jats:italic>\n                  expression and induction of sexual differentiation. The reporter lines generated allow further study of signaling pathways or screening of factors regulating sexual commitment.\n                <\/jats:p>","DOI":"10.1101\/2025.04.22.648970","type":"posted-content","created":{"date-parts":[[2025,4,24]],"date-time":"2025-04-24T00:50:18Z","timestamp":1745455818000},"source":"Crossref","is-referenced-by-count":0,"title":["A single valine to leucine switch disrupts\n                  <i>Plasmodium falciparum<\/i>\n                  AP2-G DNA binding and reveals GDV1\u2019s role in\n                  <i>ap2-g<\/i>\n                  activation"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1188-668X","authenticated-orcid":false,"given":"Surendra K.","family":"Prajapati","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, USA"},{"name":"Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-5558-2808","authenticated-orcid":false,"given":"Jeffrey X.","family":"Dong","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, USA"},{"name":"Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Belinda J.","family":"Morahan","sequence":"additional","affiliation":[{"name":"Laboratory of Malaria and Vector Research, NIAID, NIH, Rockville, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thoai","family":"Dotrang","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, USA"},{"name":"Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michelle C.","family":"Barbeau","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, USA"},{"name":"Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"April E.","family":"Williams","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7816-2948","authenticated-orcid":false,"given":"Daniel","family":"Hupalo","sequence":"additional","affiliation":[{"name":"Department of Anatomy, Physiology & Genetics, Uniformed Services University of the Health Sciences, Bethesda, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew D.","family":"Wilkerson","sequence":"additional","affiliation":[{"name":"Department of Anatomy, Physiology & Genetics, Uniformed Services University of the Health Sciences, Bethesda, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2025-8239","authenticated-orcid":false,"given":"Clifton L.","family":"Dalgard","sequence":"additional","affiliation":[{"name":"Department of Anatomy, Physiology & Genetics, Uniformed Services University of the Health Sciences, Bethesda, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0089-1096","authenticated-orcid":false,"given":"Bjorn F.C.","family":"Kafsack","sequence":"additional","affiliation":[{"name":"Department of Microbiology & Immunology, Weill Cornell Medicine, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6173-5882","authenticated-orcid":false,"given":"Manuel","family":"Llin\u00e1s","sequence":"additional","affiliation":[{"name":"Department of Biochemistry & Molecular Biology and the Huck Center for Malaria Research, Pennsylvania State University, University Park, USA"},{"name":"Department of Chemistry, Pennsylvania State University, University Park, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2749-4082","authenticated-orcid":false,"given":"Kim C.","family":"Williamson","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, USA"},{"name":"Laboratory of Malaria and Vector Research, NIAID, NIH, Rockville, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026041205111118000_2025.04.22.648970v1.1","unstructured":"WHO. WHO Reports.) (2024)."},{"key":"2026041205111118000_2025.04.22.648970v1.2","doi-asserted-by":"crossref","first-page":"e0223422","DOI":"10.1128\/spectrum.02234-22","article-title":"Plasmodium falciparum Sexual Commitment Rate Variation among Clinical Isolates and Diverse Laboratory-Adapted Lines","volume":"10","year":"2022","journal-title":"Microbiol Spectr"},{"key":"2026041205111118000_2025.04.22.648970v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-10805-w"},{"key":"2026041205111118000_2025.04.22.648970v1.4","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-19988-z"},{"key":"2026041205111118000_2025.04.22.648970v1.5","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pmed.1001788"},{"key":"2026041205111118000_2025.04.22.648970v1.6","doi-asserted-by":"crossref","unstructured":"Geiger C , et al. Declining malaria parasite prevalence and trends of asymptomatic parasitaemia in a seasonal transmission setting in North-Western Burkina Faso between 2000 and 2009-2012. Malar J 12, 27 (2013).","DOI":"10.1186\/1475-2875-12-27"},{"key":"2026041205111118000_2025.04.22.648970v1.7","doi-asserted-by":"crossref","unstructured":"Adjah J , Fiadzoe B , Ayanful-Torgby R , Amoah LE . Seasonal variations in Plasmodium falciparum genetic diversity and multiplicity of infection in asymptomatic children living in southern Ghana. BMC Infect Dis 18, 432 (2018).","DOI":"10.1186\/s12879-018-3350-z"},{"key":"2026041205111118000_2025.04.22.648970v1.8","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cix336"},{"key":"2026041205111118000_2025.04.22.648970v1.9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002964"},{"key":"2026041205111118000_2025.04.22.648970v1.10","doi-asserted-by":"publisher","DOI":"10.1038\/nature12920"},{"key":"2026041205111118000_2025.04.22.648970v1.11","doi-asserted-by":"publisher","DOI":"10.1126\/science.aan6042"},{"key":"2026041205111118000_2025.04.22.648970v1.12","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-15026-0"},{"key":"2026041205111118000_2025.04.22.648970v1.13","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0902515106"},{"key":"2026041205111118000_2025.04.22.648970v1.14","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000569"},{"key":"2026041205111118000_2025.04.22.648970v1.15","doi-asserted-by":"crossref","unstructured":"Sales-Gil R , Vagnarelli P . How HP1 Post-Translational Modifications Regulate Heterochromatin Formation and Maintenance. Cells 9, (2020).","DOI":"10.3390\/cells9061460"},{"key":"2026041205111118000_2025.04.22.648970v1.16","doi-asserted-by":"crossref","unstructured":"Papamokos GV , Kaxiras E . How to evict HP1 from H3: Using a complex salt bridge. Biophys Chem 300, 107062 (2023).","DOI":"10.1016\/j.bpc.2023.107062"},{"key":"2026041205111118000_2025.04.22.648970v1.17","doi-asserted-by":"publisher","DOI":"10.1038\/nature04254"},{"key":"2026041205111118000_2025.04.22.648970v1.18","doi-asserted-by":"publisher","DOI":"10.1038\/nature04219"},{"key":"2026041205111118000_2025.04.22.648970v1.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2011.09.004"},{"key":"2026041205111118000_2025.04.22.648970v1.20","doi-asserted-by":"crossref","unstructured":"Ganter M , et al. Plasmodium falciparum CRK4 directs continuous rounds of DNA replication during schizogony. Nat Microbiol 2, 17017 (2017).","DOI":"10.1038\/nmicrobiol.2017.38"},{"key":"2026041205111118000_2025.04.22.648970v1.21","doi-asserted-by":"publisher","DOI":"10.1021\/pr400394g"},{"key":"2026041205111118000_2025.04.22.648970v1.22","doi-asserted-by":"publisher","DOI":"10.1093\/NAR\/GKAB929"},{"key":"2026041205111118000_2025.04.22.648970v1.23","doi-asserted-by":"crossref","unstructured":"Zhang M , et al. Uncovering the essential genes of the human malaria parasite Plasmodium falciparum by saturation mutagenesis. Science 360, (2018).","DOI":"10.1126\/science.aap7847"},{"key":"2026041205111118000_2025.04.22.648970v1.24","doi-asserted-by":"crossref","unstructured":"Tiburcio M , et al. A 39-Amino-Acid C-Terminal Truncation of GDV1 Disrupts Sexual Commitment in Plasmodium falciparum. mSphere 6, (2021).","DOI":"10.1128\/mSphere.01093-20"},{"key":"2026041205111118000_2025.04.22.648970v1.25","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aaz5057"},{"key":"2026041205111118000_2025.04.22.648970v1.26","unstructured":"MalariaGen, et al. Pf7: an open dataset of Plasmodium falciparum genome variation in 20,000 worldwide samples. Wellcome Open Res 8, 22 (2023)."},{"key":"2026041205111118000_2025.04.22.648970v1.27","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-82-5-1027"},{"key":"2026041205111118000_2025.04.22.648970v1.28","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-02460-2"},{"key":"2026041205111118000_2025.04.22.648970v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2017.05.006"},{"key":"2026041205111118000_2025.04.22.648970v1.30","doi-asserted-by":"publisher","DOI":"10.1093\/NAR\/GKAB683"},{"key":"2026041205111118000_2025.04.22.648970v1.31","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-023-01497-6"},{"key":"2026041205111118000_2025.04.22.648970v1.32","doi-asserted-by":"crossref","unstructured":"Yuda M , Kaneko I , Murata Y , Iwanaga S , Nishi T . Mechanisms of triggering malaria gametocytogenesis by AP2-G. Parasitol Int 84, 102403 (2021).","DOI":"10.1016\/j.parint.2021.102403"},{"key":"2026041205111118000_2025.04.22.648970v1.33","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-018-0223-6"},{"key":"2026041205111118000_2025.04.22.648970v1.34","doi-asserted-by":"publisher","DOI":"10.1038\/nature19804"},{"key":"2026041205111118000_2025.04.22.648970v1.35","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01144-06"},{"key":"2026041205111118000_2025.04.22.648970v1.36","doi-asserted-by":"crossref","unstructured":"Jillapalli R , et al. A Plasmodium berghei putative serine-threonine kinase 2 (PBANKA_0311400) is required for late liver stage development and timely initiation of blood stage infection. Biol Open 8, (2019).","DOI":"10.1242\/bio.042028"},{"key":"2026041205111118000_2025.04.22.648970v1.37","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1217712110"},{"key":"2026041205111118000_2025.04.22.648970v1.38","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.2c00607"},{"key":"2026041205111118000_2025.04.22.648970v1.39","doi-asserted-by":"crossref","unstructured":"Antonova-Koch Y , et al. Open-source discovery of chemical leads for next-generation chemoprotective antimalarials. Science 362, (2018).","DOI":"10.1126\/science.aat9446"},{"key":"2026041205111118000_2025.04.22.648970v1.40","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1011517"},{"key":"2026041205111118000_2025.04.22.648970v1.41","doi-asserted-by":"publisher","DOI":"10.1021\/jacsau.4c00674"},{"key":"2026041205111118000_2025.04.22.648970v1.42","doi-asserted-by":"crossref","unstructured":"Prajapati SK , Joshi H , Dev V , Dua VK . Molecular epidemiology of Plasmodium vivax anti-folate resistance in India. Malar J 10, 102 (2011).","DOI":"10.1186\/1475-2875-10-102"},{"key":"2026041205111118000_2025.04.22.648970v1.43","doi-asserted-by":"crossref","unstructured":"Beckmann JF , Van Vaerenberghe K , Akwa DE , Cooper BS . A single mutation weakens symbiont-induced reproductive manipulation through reductions in deubiquitylation efficiency. Proc Natl Acad Sci U S A 118, (2021).","DOI":"10.1073\/pnas.2113271118"},{"key":"2026041205111118000_2025.04.22.648970v1.44","doi-asserted-by":"publisher","DOI":"10.1210\/me.7.1.37"},{"key":"2026041205111118000_2025.04.22.648970v1.45","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopsych.2006.03.017"},{"key":"2026041205111118000_2025.04.22.648970v1.46","doi-asserted-by":"crossref","unstructured":"Rajebhosale P , et al. Neuregulin1 nuclear signaling influences adult neurogenesis and regulates a schizophrenia susceptibility gene network within the mouse dentate gyrus. bioRxiv, 2022.2008.2010.503469 (2024).","DOI":"10.1101\/2022.08.10.503469"},{"key":"2026041205111118000_2025.04.22.648970v1.47","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-0716-2189-9_17"},{"key":"2026041205111118000_2025.04.22.648970v1.48","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2925"},{"key":"2026041205111118000_2025.04.22.648970v1.49","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2812"},{"key":"2026041205111118000_2025.04.22.648970v1.50","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks596"},{"key":"2026041205111118000_2025.04.22.648970v1.51","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btp324"},{"key":"2026041205111118000_2025.04.22.648970v1.52","doi-asserted-by":"publisher","DOI":"10.1038\/ng.806"},{"key":"2026041205111118000_2025.04.22.648970v1.53","unstructured":"https:\/\/www.genome.jp\/tools\/motif\/."},{"key":"2026041205111118000_2025.04.22.648970v1.54","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1001165"},{"key":"2026041205111118000_2025.04.22.648970v1.55","unstructured":"One-way ANOVA followed by Dunnett\u2019s multiple comparisons test was performed using GraphPad Prism version 10.0.0 for Windows, GraphPad Software, Boston, Massachusetts USA, www.graphpad.com."},{"key":"2026041205111118000_2025.04.22.648970v1.56","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4074"},{"key":"2026041205111118000_2025.04.22.648970v1.57","doi-asserted-by":"crossref","unstructured":"Anna O. Chertkova MM , Marten Postma , Nikki van Bommel , Sanne van der Niet , Kevin L. Batenburg , Linda Joosen , Theodorus W.J. Gadella Jr. , Yasushi Okada , Joachim Goedhart . Robust and Bright Genetically Encoded Fluorescent Markers for Highlighting Structures and Compartments in Mammalian Cells. bioRxiv (2017).","DOI":"10.1101\/160374"},{"key":"2026041205111118000_2025.04.22.648970v1.58","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4222"},{"key":"2026041205111118000_2025.04.22.648970v1.59","doi-asserted-by":"publisher","DOI":"10.1038\/nature12970"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.04.22.648970","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:04:14Z","timestamp":1782277454000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.04.22.648970"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,23]]},"references-count":59,"URL":"https:\/\/doi.org\/10.1101\/2025.04.22.648970","relation":{"is-preprint-of":[{"id-type":"doi","id":"10.1038\/s41467-026-68416-1","asserted-by":"subject"}]},"subject":[],"published":{"date-parts":[[2025,4,23]]},"subtype":"preprint"}}