{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T19:05:36Z","timestamp":1772132736763,"version":"3.50.1"},"reference-count":21,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T00:00:00Z","timestamp":1772064000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science and Technology of Japan","doi-asserted-by":"crossref","award":["24K23322"],"award-info":[{"award-number":["24K23322"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science and Technology of Japan","doi-asserted-by":"crossref","award":["18H04675, 21K06992 and 25K02488"],"award-info":[{"award-number":["18H04675, 21K06992 and 25K02488"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science and Technology of Japan","doi-asserted-by":"crossref","award":["19K07627 and 24K10273"],"award-info":[{"award-number":["19K07627 and 24K10273"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"crossref"}]},{"name":"AMED Japan Program for Infectious Diseases Research and Infrastructure","award":["23wm0325036 and 25wm0325071"],"award-info":[{"award-number":["23wm0325036 and 25wm0325071"]}]},{"name":"AMED Japan Program for Infectious Diseases Research and Infrastructure","award":["23wm0325036 and 25wm0325071"],"award-info":[{"award-number":["23wm0325036 and 25wm0325071"]}]},{"name":"Kaketsuken Foundation"},{"DOI":"10.13039\/501100008971","name":"Nagase Science Technology Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100008971","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016925","name":"Takeda Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100016925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007428","name":"Naito Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007428","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>\n                    <jats:italic>Entamoeba histolytica<\/jats:italic>\n                    , a protozoan parasite, causes amebiasis. Amebiasis is mainly transmitted by oral ingestion of cysts. Cysts are produced in the large intestine of humans from proliferative trophozoites by a cell differentiation process called encystation. The\n                    <jats:italic>Entamoeba<\/jats:italic>\n                    cyst wall consists of chitins and proteins that include chitinase and Jacob and Jessie lectins. During encystation, these components are synthesized and layered around encysting\n                    <jats:italic>Entamoeba<\/jats:italic>\n                    cells. The structures of these components are well studied; however, the detailed timings of their synthesis (the transcription of the encoding genes and the translation of the resulting mRNAs) and of changes in their localization during encystation are poorly understood. Here, we performed quantitative RT-PCR and an approach combining western blotting and immunofluorescence, confocal, and immunoelectron microscopy to analyze\n                    <jats:italic>Entamoeba invadens<\/jats:italic>\n                    cells that were sampled with short-time intervals during encystation. A chitinase inhibitor, D-B-09, which disrupts the compression of chitin fibers was used to analyze component interaction with chitin fibers. All genes encoding cyst wall proteins were stage-specifically transcribed and translated, and post-translationally modified forms of Jacob1\/3 were trafficked to the cyst wall before Jessie3a and Chitinase1\/4 were simultaneously localized in the cyst wall. The trafficking of Jacob lectins to the cyst wall and their co-localization with chitin fibers in encysting cells were not affected by D-B-09, while the localization of Jessie protein was impaired, indicating that localizations of Jacob and Jessie lectins are spatially different positions via different modes in cyst wall. These results indicate that cyst wall components are functionally linked and that they play different roles during\n                    <jats:italic>Entamoeba<\/jats:italic>\n                    cyst wall formation. Immunoelectron microscopy confirmed the immunofluorescence and confocal microscopy results. Importantly, immunoelectron microscopy also indicated that the\n                    <jats:italic>Entamoeba<\/jats:italic>\n                    cyst wall consists of a biphasic structure of electron-light (inner) and electron-dense (outer) areas.\n                  <\/jats:p>","DOI":"10.1371\/journal.ppat.1013940","type":"journal-article","created":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T18:21:29Z","timestamp":1772130089000},"page":"e1013940","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Trafficking and organization of cyst wall components into a robust biphasic structure in Entamoeba"],"prefix":"10.1371","volume":"22","author":[{"given":"Tam Kha","family":"Vo","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroki","family":"Yoshida","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7042-2266","authenticated-orcid":true,"given":"Fumika","family":"Mi-ichi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,2,26]]},"reference":[{"issue":"10","key":"ppat.1013940.ref001","doi-asserted-by":"crossref","first-page":"1134","DOI":"10.1016\/j.jiph.2022.08.013","article-title":"Updates on the worldwide burden of amoebiasis: A case series and literature review","volume":"15","author":"J Nasrallah","year":"2022","journal-title":"J Infect Public Health"},{"issue":"2","key":"ppat.1013940.ref002","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1111\/mmi.13131","article-title":"Molecular biology research to benefit patients with Entamoeba histolytica infection","volume":"98","author":"K Watanabe","year":"2015","journal-title":"Mol Microbiol"},{"issue":"16","key":"ppat.1013940.ref003","doi-asserted-by":"crossref","first-page":"1565","DOI":"10.1056\/NEJMra022710","article-title":"Amebiasis","volume":"348","author":"R Haque","year":"2003","journal-title":"N Engl J Med"},{"key":"ppat.1013940.ref004","doi-asserted-by":"crossref","first-page":"102844","DOI":"10.1016\/j.parint.2023.102844","article-title":"Links between cholesteryl sulfate-dependent and -independent processes in the morphological and physiological changes of Entamoeba encystation","volume":"99","author":"F Mi-Ichi","year":"2024","journal-title":"Parasitol Int"},{"issue":"10","key":"ppat.1013940.ref005","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1005845","article-title":"Entamoeba Encystation: New Targets to Prevent the Transmission of Amebiasis","volume":"12","author":"F Mi-Ichi","year":"2016","journal-title":"PLoS Pathog"},{"issue":"4","key":"ppat.1013940.ref006","doi-asserted-by":"crossref","first-page":"873","DOI":"10.3390\/microorganisms9040873","article-title":"Encystation of Entamoeba histolytica in Axenic Culture","volume":"9","author":"J Wesel","year":"2021","journal-title":"Microorganisms"},{"issue":"1","key":"ppat.1013940.ref007","doi-asserted-by":"crossref","first-page":"20","DOI":"10.3390\/pathogens13010020","article-title":"Novel Insights into the Wattle and Daub Model of Entamoeba Cyst Wall Formation and the Importance of Actin Cytoskeleton","volume":"13","author":"D Krishnan","year":"2023","journal-title":"Pathogens"},{"issue":"7","key":"ppat.1013940.ref008","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1000498","article-title":"Evidence for a \u201cwattle and daub\u201d model of the cyst wall of entamoeba","volume":"5","author":"A Chatterjee","year":"2009","journal-title":"PLoS Pathog"},{"issue":"5","key":"ppat.1013940.ref009","doi-asserted-by":"crossref","first-page":"836","DOI":"10.1128\/EC.5.5.836-848.2006","article-title":"Unique posttranslational modifications of chitin-binding lectins of Entamoeba invadens cyst walls","volume":"5","author":"KL Van Dellen","year":"2006","journal-title":"Eukaryot Cell"},{"issue":"1","key":"ppat.1013940.ref010","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.pt.2010.09.002","article-title":"A simple fibril and lectin model for cyst walls of Entamoeba and perhaps Giardia","volume":"27","author":"J Samuelson","year":"2011","journal-title":"Trends Parasitol"},{"key":"ppat.1013940.ref011","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1007\/978-981-13-7318-3_11","article-title":"Human Chitinases: Structure, Function, and Inhibitor Discovery","volume":"1142","author":"A Kumar","year":"2019","journal-title":"Adv Exp Med Biol"},{"issue":"1","key":"ppat.1013940.ref012","doi-asserted-by":"crossref","DOI":"10.1128\/Spectrum.00511-21","article-title":"Entamoeba Chitinase is Required for Mature Round Cyst Formation","volume":"9","author":"F Mi-Ichi","year":"2021","journal-title":"Microbiol Spectr"},{"issue":"6","key":"ppat.1013940.ref013","doi-asserted-by":"crossref","first-page":"3259","DOI":"10.1128\/IAI.70.6.3259-3263.2002","article-title":"Entamoeba histolytica lectins contain unique 6-Cys or 8-Cys chitin-binding domains","volume":"70","author":"K Van Dellen","year":"2002","journal-title":"Infect Immun"},{"issue":"7","key":"ppat.1013940.ref014","doi-asserted-by":"crossref","DOI":"10.1186\/gb-2013-14-7-r77","article-title":"The genome and transcriptome of the enteric parasite Entamoeba invadens, a model for encystation","volume":"14","author":"GM Ehrenkaufer","year":"2013","journal-title":"Genome Biol"},{"issue":"9","key":"ppat.1013940.ref015","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0074840","article-title":"Transcriptome analysis of encystation in Entamoeba invadens","volume":"8","author":"AE De C\u00e1diz","year":"2013","journal-title":"PLoS One"},{"issue":"12","key":"ppat.1013940.ref016","doi-asserted-by":"crossref","first-page":"1305","DOI":"10.1017\/S0031182020001079","article-title":"The dynamics of ultrastructural changes during Entamoeba invadens encystation","volume":"147","author":"EAA Mousa","year":"2020","journal-title":"Parasitology"},{"issue":"7","key":"ppat.1013940.ref017","doi-asserted-by":"crossref","first-page":"4217","DOI":"10.1128\/IAI.68.7.4217-4224.2000","article-title":"The most abundant glycoprotein of amebic cyst walls (Jacob) is a lectin with five Cys-rich, chitin-binding domains","volume":"68","author":"M Frisardi","year":"2000","journal-title":"Infect Immun"},{"issue":"3","key":"ppat.1013940.ref018","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1078\/1434461041844204","article-title":"Characterization of chitin synthases from Entamoeba","volume":"155","author":"E Campos-G\u00f3ngora","year":"2004","journal-title":"Protist"},{"issue":"7931","key":"ppat.1013940.ref019","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1038\/s41586-022-05244-5","article-title":"Structural basis for directional chitin biosynthesis","volume":"610","author":"W Chen","year":"2022","journal-title":"Nature"},{"key":"ppat.1013940.ref020","doi-asserted-by":"crossref","first-page":"250","DOI":"10.3389\/fcimb.2018.00250","article-title":"A Flow Cytometry Method for Dissecting the Cell Differentiation Process of Entamoeba Encystation","volume":"8","author":"F Mi-Ichi","year":"2018","journal-title":"Front Cell Infect Microbiol"},{"issue":"2","key":"ppat.1013940.ref021","doi-asserted-by":"crossref","DOI":"10.1128\/mSphere.00174-21","article-title":"Stage-Specific De Novo Synthesis of Very-Long-Chain Dihydroceramides Confers Dormancy to Entamoeba Parasites","volume":"6","author":"F Mi-Ichi","year":"2021","journal-title":"mSphere"}],"container-title":["PLOS Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.ppat.1013940","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T18:21:36Z","timestamp":1772130096000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.ppat.1013940"}},"subtitle":[],"editor":[{"given":"Tracey J.","family":"Lamb","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2026,2,26]]},"references-count":21,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2,26]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.ppat.1013940","relation":{},"ISSN":["1553-7374"],"issn-type":[{"value":"1553-7374","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,26]]}}}