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In\n              <jats:italic toggle=\"yes\">Saccharomyces cerevisiae,<\/jats:italic>\n              synthesis of inositol phosphates occurs through the phosphoinositide phospholipase C (PLC)-catalyzed hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP\n              <jats:sub>2<\/jats:sub>\n              ) into inositol 1,4,5-trisphosphate (IP\n              <jats:sub>3<\/jats:sub>\n              ) and diacylglycerol and further IP\n              <jats:sub>3<\/jats:sub>\n              phosphorylation by additional kinases that leads to the formation of highly phosphorylated inositol derivatives, known as inositol pyrophosphates. Inositol-tetrakisphosphate 1-kinase (ITPK1) is an enzyme that mediates a PLC-independent inositol polyphosphate synthesis through phosphorylation of inositol monophosphates and other intermediates in the cytosol. In this work, we identified and characterized a\n              <jats:italic toggle=\"yes\">Trypanosoma cruzi<\/jats:italic>\n              ITPK1 (TcITPK1) homolog. The ability of TcITPK1 to act as the mediator for this alternative pathway was established through\n              <jats:italic toggle=\"yes\">plc1<\/jats:italic>\n              \u0394 and\n              <jats:italic toggle=\"yes\">plc1<\/jats:italic>\n              \u0394\n              <jats:italic toggle=\"yes\">isc1<\/jats:italic>\n              \u0394 yeast complementation assays and SAX-HPLC analyses of radioactively labeled inositol. TcITPK1 localizes to the cytosol, and knockout attempts of\n              <jats:italic toggle=\"yes\">TcITPK1<\/jats:italic>\n              revealed that only one allele was replaced by the DNA donor cassette at the specific locus, suggesting that\n              <jats:italic toggle=\"yes\">null<\/jats:italic>\n              alleles may have lethal effects in epimastigotes. Ablation of\n              <jats:italic toggle=\"yes\">T. cruzi<\/jats:italic>\n              phosphoinositide phospholipase C 1 (\n              <jats:italic toggle=\"yes\">TcPI-PLC1<\/jats:italic>\n              ) affected the synthesis of IP\n              <jats:sub>3<\/jats:sub>\n              from glucose 6-phosphate but did not affect the synthesis of inositol polyphosphates, while ablation of inositol phosphosphingolipid phospholipase (\n              <jats:italic toggle=\"yes\">TcISC1<\/jats:italic>\n              ) affected the synthesis of inositol polyphosphates, thus revealing that the PLC-independent pathway using either glucose 6-phosphate or inositol phosphoceramide is involved in the synthesis of inositol polyphosphates, while the PLC-dependent pathway is involved in IP\n              <jats:sub>3<\/jats:sub>\n              formation needed for Ca\n              <jats:sup>2+<\/jats:sup>\n              signaling.\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>\n              Millions of people are infected with\n              <jats:italic toggle=\"yes\">Trypanosoma cruzi,<\/jats:italic>\n              and the current treatment is not satisfactory. Inositol pyrophosphates have been established as important signaling molecules. Our work demonstrates the presence of a phospholipase C-independent pathway for the synthesis of inositol pyrophosphates in\n              <jats:italic toggle=\"yes\">T. cruzi<\/jats:italic>\n              . Furthermore, we demonstrate that this pathway starts with the synthesis of inositol monophosphates from glucose 6-phosphate or from inositol phosphoceramide, linking it to carbohydrate and sphingolipid metabolism. The essentiality of the pathway for the survival of\n              <jats:italic toggle=\"yes\">T. cruzi<\/jats:italic>\n              infective stages makes it an ideal drug target for treating American trypanosomiasis.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/mbio.03318-24","type":"journal-article","created":{"date-parts":[[2025,4,4]],"date-time":"2025-04-04T05:29:22Z","timestamp":1743744562000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":3,"title":["Generation of inositol polyphosphates through a phospholipase C-independent pathway involving carbohydrate and sphingolipid metabolism in\n            <i>Trypanosoma cruzi<\/i>"],"prefix":"10.1128","volume":"16","author":[{"given":"Mayara S.","family":"Bertolini","sequence":"first","affiliation":[{"name":"Department of Cellular Biology, Center for Tropical and Emerging Global Diseases, University of Georgia","place":["Athens, 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