{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T18:37:02Z","timestamp":1793126222999,"version":"4.5.0"},"reference-count":53,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2014,5,1]],"date-time":"2014-05-01T00:00:00Z","timestamp":1398902400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Eukaryot Cell"],"published-print":{"date-parts":[[2014,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Human African trypanosomiasis (HAT) is a debilitating and fatal vector-borne disease. Polyamine biosynthesis is the target of one of the key drugs (eflornithine) used for the treatment of late-stage disease, suggesting that the pathway might be exploited for the identification of additional drug targets. The polyamine spermidine is required in trypanosomatid parasites for formation of a unique redox cofactor termed trypanothione, which is formed from the conjugation of glutathione to spermidine. Here we characterize recombinant\n            <jats:named-content content-type=\"genus-species\">Trypanosoma brucei<\/jats:named-content>\n            glutathione synthetase (\n            <jats:italic>Tb<\/jats:italic>\n            GS) and show that depletion of\n            <jats:italic>Tb<\/jats:italic>\n            GS in blood-form parasites using a regulated knockout strategy leads to loss of trypanothione and to cell death as quantified by fluorescence-activated cell sorter (FACS) analysis. These data suggest that &gt;97% depletion of\n            <jats:italic>Tb<\/jats:italic>\n            GS is required before trypanothione is depleted and cell growth arrest is observed. Exogenous glutathione was able to partially compensate for the loss of\n            <jats:italic>Tb<\/jats:italic>\n            GS, suggesting that parasites are able to transport intact glutathione. Finally, reduced expression of\n            <jats:italic>Tb<\/jats:italic>\n            GS leads to increased levels of upstream glutathione biosynthetic enzymes and decreased expression of polyamine biosynthetic enzymes, providing evidence that the cells cross regulate the two branches of the trypanothione biosynthetic pathway to maintain spermidine and trypanothione homeostasis.\n          <\/jats:p>","DOI":"10.1128\/ec.00015-14","type":"journal-article","created":{"date-parts":[[2014,3,8]],"date-time":"2014-03-08T05:39:07Z","timestamp":1394257147000},"page":"614-624","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":15,"title":["Genetic Validation of Trypanosoma brucei Glutathione Synthetase as an Essential Enzyme"],"prefix":"10.1128","volume":"13","author":[{"given":"Chelsea","family":"Pratt","sequence":"first","affiliation":[{"name":"Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suong","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Margaret A.","family":"Phillips","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI33945"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(12)70296-X"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.2174\/156802611795429167"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1097\/QCO.0b013e32833f9fd0"},{"key":"e_1_3_3_6_2","volume-title":"Human African trypanosomiasis","author":"World Health Organization","year":"2012","unstructured":"World Health Organization. January 2012, posting date . Human African trypanosomiasis. World Health Organization, Geneva, Switzerland. http:\/\/www.who.int\/trypanosomiasis_african\/en\/ 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