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The virulence of<jats:named-content content-type=\"genus-species\">E. histolytica<\/jats:named-content>correlates with the degree of host cell engulfment, or phagocytosis, and<jats:named-content content-type=\"genus-species\">E. histolytica<\/jats:named-content>phagocytosis alters amebic gene expression in a feed-forward manner that results in an increased phagocytic ability. Here, we used a streamlined RNA interference screen to silence the expression of 15 genes whose expression was upregulated in phagocytic<jats:named-content content-type=\"genus-species\">E. histolytica<\/jats:named-content>trophozoites to determine whether these genes actually function in the phagocytic process. When five of these genes were silenced, amebic strains with significant decreases in the ability to phagocytose apoptotic host cells were produced. Phagocytosis of live host cells, however, was largely unchanged, and the defects were surprisingly specific for phagocytosis. Two of the five encoded proteins, which we named<jats:named-content content-type=\"genus-species\">E. histolytica<\/jats:named-content>ILWEQ (EhILWEQ) and<jats:named-content content-type=\"genus-species\">E. histolytica<\/jats:named-content>BAR (EhBAR), were chosen for localization via SNAP tag labeling and localized to the site of partially formed phagosomes. Therefore, both EhILWEQ and EhBAR appear to contribute to<jats:named-content content-type=\"genus-species\">E. histolytica<\/jats:named-content>virulence through their function in phagocytosis, and the large proportion (5\/15 [33%]) of gene-silenced strains with a reduced ability to phagocytose host cells validates the previously published microarray data set demonstrating feed-forward control of<jats:named-content content-type=\"genus-species\">E. histolytica<\/jats:named-content>phagocytosis. Finally, although only limited conclusions can be drawn from studies using the virulence-deficient G3<jats:named-content content-type=\"genus-species\">Entamoeba<\/jats:named-content>strain, the relative specificity of the defects induced for phagocytosis of apoptotic cells but not healthy cells suggests that cell killing may play a rate-limiting role in the process of<jats:named-content content-type=\"genus-species\">Entamoeba histolytica<\/jats:named-content>host cell engulfment.<\/jats:p>","DOI":"10.1128\/iai.01325-15","type":"journal-article","created":{"date-parts":[[2016,1,26]],"date-time":"2016-01-26T03:48:57Z","timestamp":1453780137000},"page":"1045-1053","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":9,"title":["Knockdown of Five Genes Encoding Uncharacterized Proteins Inhibits Entamoeba histolytica Phagocytosis of Dead Host Cells"],"prefix":"10.1128","volume":"84","author":[{"given":"Adam","family":"Sateriale","sequence":"first","affiliation":[{"name":"Department of Medicine, University of Vermont College of Medicine, Burlington, Vermont, USA"},{"name":"Cellular, Molecular, and Biomedical Sciences Program, University of Vermont College of Medicine, Burlington, Vermont, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Miller","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Vermont College of Medicine, Burlington, Vermont, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher D.","family":"Huston","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Vermont College of Medicine, Burlington, Vermont, USA"},{"name":"Cellular, Molecular, and Biomedical Sciences Program, University of Vermont College of Medicine, Burlington, Vermont, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","first-page":"97","article-title":"Amoebiasis","volume":"72","author":"WHO","year":"1997","unstructured":"WHO. 1997. 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A hidden Markov model for predicting transmembrane helices in protein sequences. Proc Int Conf Intell Syst Mol Biol 6:175\u2013182.","journal-title":"Proc Int Conf Intell Syst Mol Biol"}],"container-title":["Infection and Immunity"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/IAI.01325-15","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/IAI.01325-15","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,6,13]],"date-time":"2024-06-13T23:45:29Z","timestamp":1718322329000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/IAI.01325-15"}},"subtitle":[],"editor":[{"given":"J. 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