{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T14:00:42Z","timestamp":1785765642794,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,30]]},"group-title":"Cell Biology","reference-count":35,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T00:00:00Z","timestamp":1785369600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"award":["N\/A"],"award-info":[{"award-number":["N\/A"]}],"id":[{"id":"https:\/\/ror.org\/0405mnx93","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R21AI156225"],"award-info":[{"award-number":["R21AI156225"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The eukaryotic pathogen\n                  <jats:italic>Leishmania major<\/jats:italic>\n                  causes disfiguring cutaneous lesions, whose resolution can be complicated by secondary bacterial infections. Bacteria, including\n                  <jats:italic>Aeromonas<\/jats:italic>\n                  spp., also interact with\n                  <jats:italic>L. major<\/jats:italic>\n                  promastigotes in the sandfly midgut. The mechanisms by which\n                  <jats:italic>L. major<\/jats:italic>\n                  competes with bacteria and resists their toxins are poorly defined. Prior work proposed that\n                  <jats:italic>L. major<\/jats:italic>\n                  resists the\n                  <jats:italic>Aeromonas<\/jats:italic>\n                  -produced pore-forming toxin aerolysin using an altered GPI-anchor. However, we found that\n                  <jats:italic>L. major<\/jats:italic>\n                  is sensitive to aerolysin. Here, we determined the mechanism by which\n                  <jats:italic>L. major<\/jats:italic>\n                  promastigotes are sensitive to aerolysin, using flow cytometry and biochemical approaches to analyze promastigotes genetically deficient in enzymes that produce key membrane components. The virulence factor lipophosphoglycan protected\n                  <jats:italic>L. major<\/jats:italic>\n                  from aerolysin cytotoxicity. The metalloproteinase GP63 exerted the necessary furin-like protease activity to activate aerolysin. Leishmanial GPI-anchored proteins were necessary for aerolysin heptamerization and killing of\n                  <jats:italic>L. major<\/jats:italic>\n                  promastigotes. Finally, mutation of the GPI-anchor binding domain of aerolysin crippled its cytotoxicity, consistent with its reliance on the GPI-anchor binding site to engage GPI anchors on the surface of\n                  <jats:italic>L. major<\/jats:italic>\n                  promastigotes. Taken together, we propose the\n                  <jats:italic>L. major<\/jats:italic>\n                  virulence factor lipophosphoglycan defends against pore-forming toxins made by bacterial competitors, while the GP63 metalloproteinase activates pro-aerolysin like furin. Overall, this study highlights approaches microbes use to compete with each other.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Graphical Abstract<\/jats:title>\n                  <jats:p>\n                    <jats:bold>\n                      Aerolysin cytotoxicity depends on gp63 and LPG in\n                      <jats:italic>Leishmania major<\/jats:italic>\n                      promastigotes.\n                    <\/jats:bold>\n                    (A) Wild type\n                    <jats:italic>Leishmania major<\/jats:italic>\n                    promastigotes are sensitive to aerolysin, which forms lethal heptameric pore complexes in the plasma membrane (B)\n                    <jats:italic>\n                      L. major lpg1\n                      <jats:sup>\u2014<\/jats:sup>\n                    <\/jats:italic>\n                    promastigotes are highly sensitive to aerolysin challenge because they lack LPG. (C)\n                    <jats:italic>\n                      L. major gp63\n                      <jats:sup>\u2014<\/jats:sup>\n                    <\/jats:italic>\n                    promastigotes have wild type sensitivity to aerolysin challenge but resist pro-aerolysin. (D)\n                    <jats:italic>\n                      L. major gpi8\n                      <jats:sup>\u2014<\/jats:sup>\n                    <\/jats:italic>\n                    knockout promastigotes are resistant to aerolysin and show no heptameric pore complexes in the plasma membrane. Created in BioRender.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.07.29.741567","type":"posted-content","created":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:35:14Z","timestamp":1785425714000},"source":"Crossref","is-referenced-by-count":0,"title":["Leishmanial GP63 acts as a protease for the small pore forming toxin aerolysin"],"prefix":"10.64898","author":[{"given":"Chaitanya S.","family":"Haram","sequence":"first","affiliation":[{"name":"Department of Biological Sciences Texas Tech University Lubbock, TX 79409"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sebastian","family":"Salinas","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences Texas Tech University Lubbock, TX 79409"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Salma Waheed","family":"Sheikh","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences Texas Tech University Lubbock, TX 79409"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences Texas Tech University Lubbock, TX 79409"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8637-8538","authenticated-orcid":false,"given":"Peter A.","family":"Keyel","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences Texas Tech University Lubbock, TX 79409"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026080306201669000_2026.07.29.741567v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-025-00663-w"},{"key":"2026080306201669000_2026.07.29.741567v1.2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0010609"},{"key":"2026080306201669000_2026.07.29.741567v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-02388-8"},{"key":"2026080306201669000_2026.07.29.741567v1.4","doi-asserted-by":"publisher","DOI":"10.3390\/ijerph14070777"},{"key":"2026080306201669000_2026.07.29.741567v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2023.104745"},{"key":"2026080306201669000_2026.07.29.741567v1.6","doi-asserted-by":"publisher","DOI":"10.1128\/jb.171.5.2523-2527.1989"},{"key":"2026080306201669000_2026.07.29.741567v1.7","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.49.32656"},{"key":"2026080306201669000_2026.07.29.741567v1.8","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.32.22604"},{"key":"2026080306201669000_2026.07.29.741567v1.9","doi-asserted-by":"publisher","DOI":"10.1046\/j.1462-5822.2000.00037.x"},{"key":"2026080306201669000_2026.07.29.741567v1.10","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1010640"},{"key":"2026080306201669000_2026.07.29.741567v1.11","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.11.4.1183"},{"key":"2026080306201669000_2026.07.29.741567v1.12","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.4.2355"},{"key":"2026080306201669000_2026.07.29.741567v1.13","doi-asserted-by":"publisher","DOI":"10.1021\/bi00151a026"},{"key":"2026080306201669000_2026.07.29.741567v1.14","doi-asserted-by":"publisher","DOI":"10.3791\/64341"},{"key":"2026080306201669000_2026.07.29.741567v1.15","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.076182"},{"key":"2026080306201669000_2026.07.29.741567v1.16","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abl6367"},{"key":"2026080306201669000_2026.07.29.741567v1.17","doi-asserted-by":"publisher","DOI":"10.3390\/toxins18010046"},{"key":"2026080306201669000_2026.07.29.741567v1.18","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.14139"},{"key":"2026080306201669000_2026.07.29.741567v1.19","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1004427"},{"key":"2026080306201669000_2026.07.29.741567v1.20","doi-asserted-by":"publisher","DOI":"10.3390\/ijms26178473"},{"key":"2026080306201669000_2026.07.29.741567v1.21","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.140.3.525"},{"key":"2026080306201669000_2026.07.29.741567v1.22","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.160257897"},{"key":"2026080306201669000_2026.07.29.741567v1.23","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-6851(01)00432-7"},{"key":"2026080306201669000_2026.07.29.741567v1.24","doi-asserted-by":"publisher","DOI":"10.21203\/rs.21203.rs-10145497\/v10145491"},{"key":"2026080306201669000_2026.07.29.741567v1.25","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/cdg584"},{"key":"2026080306201669000_2026.07.29.741567v1.26","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19970320)53:6<594::AID-BIT7>3.0.CO;2-H"},{"key":"2026080306201669000_2026.07.29.741567v1.27","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(02)02274-3"},{"key":"2026080306201669000_2026.07.29.741567v1.28","doi-asserted-by":"crossref","first-page":"16396","DOI":"10.1016\/S0021-9258(18)42016-9","article-title":"Human furin is a calcium-dependent serine endoprotease that recognizes the sequence Arg-X-X-Arg and efficiently cleaves anthrax toxin protective antigen","volume":"267","year":"1992","journal-title":"J Biol Chem"},{"key":"2026080306201669000_2026.07.29.741567v1.29","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.36.25411"},{"key":"2026080306201669000_2026.07.29.741567v1.30","doi-asserted-by":"publisher","DOI":"10.1038\/nsb839"},{"key":"2026080306201669000_2026.07.29.741567v1.31","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.261018"},{"key":"2026080306201669000_2026.07.29.741567v1.32","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182000056304"},{"key":"2026080306201669000_2026.07.29.741567v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/0166-6851(91)90007-S"},{"key":"2026080306201669000_2026.07.29.741567v1.34","doi-asserted-by":"publisher","DOI":"10.3791\/4227"},{"key":"2026080306201669000_2026.07.29.741567v1.35","doi-asserted-by":"publisher","DOI":"10.1038\/cdd.2017.11"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.07.29.741567","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T13:20:25Z","timestamp":1785763225000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.07.29.741567"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,30]]},"references-count":35,"URL":"https:\/\/doi.org\/10.64898\/2026.07.29.741567","relation":{},"subject":[],"published":{"date-parts":[[2026,7,30]]},"subtype":"preprint"}}