{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T05:21:05Z","timestamp":1791091265805,"version":"4.1.0"},"reference-count":43,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2014,6,1]],"date-time":"2014-06-01T00:00:00Z","timestamp":1401580800000},"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":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2014,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Previously, through a proteomic analysis, proliferating cell nuclear antigen (PCNA) was found to be overexpressed in the sodium antimony gluconate (SAG)-resistant clinical isolate compared to that in the SAG-sensitive clinical isolate of<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Leishmania donovani<\/jats:named-content>. The present study was designed to explore the potential role of the PCNA protein in SAG resistance in<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">L. donovani<\/jats:named-content>. For this purpose, the protein was cloned, overexpressed, purified, and modeled. Western blot (WB) and real-time PCR (RT-PCR) analyses confirmed that PCNA was overexpressed by \u22653-fold in the log phase, stationary phase, and peanut agglutinin isolated procyclic and metacyclic stages of the promastigote form and by \u223c5-fold in the amastigote form of the SAG-resistant isolate compared to that in the SAG-sensitive isolate.<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">L. donovani<\/jats:named-content>PCNA (LdPCNA) was overexpressed as a green fluorescent protein (GFP) fusion protein in a SAG-sensitive clinical isolate of<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">L. donovani<\/jats:named-content>, and modulation of the sensitivities of the transfectants to pentavalent antimonial (Sb<jats:sup>V<\/jats:sup>) and trivalent antimonial (Sb<jats:sup>III<\/jats:sup>) drugs was assessed<jats:italic>in vitro<\/jats:italic>against promastigotes and intracellular (J774A.1 cell line) amastigotes, respectively. Overexpression of LdPCNA in the SAG-sensitive isolate resulted in an increase in the 50% inhibitory concentrations (IC<jats:sub>50<\/jats:sub>) of Sb<jats:sup>V<\/jats:sup>(from 41.2 \u00b1 0.6 \u03bcg\/ml to 66.5 \u00b1 3.9 \u03bcg\/ml) and Sb<jats:sup>III<\/jats:sup>(from 24.0 \u00b1 0.3 \u03bcg\/ml to 43.4 \u00b1 1.8 \u03bcg\/ml). Moreover, PCNA-overexpressing promastigote transfectants exhibited less DNA fragmentation compared to that of wild-type SAG-sensitive parasites upon Sb<jats:sup>III<\/jats:sup>treatment. In addition, SAG-induced nitric oxide (NO) production was found to be significantly inhibited in the macrophages infected with the transfectants compared with that in wild-type SAG-sensitive parasites. Consequently, we infer that LdPCNA has a significant role in SAG resistance in<jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">L. donovani<\/jats:named-content>clinical isolates, which warrants detailed investigations regarding its mechanism.<\/jats:p>","DOI":"10.1128\/aac.01847-13","type":"journal-article","created":{"date-parts":[[2014,3,11]],"date-time":"2014-03-11T03:54:04Z","timestamp":1394510044000},"page":"2997-3007","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":19,"title":["Characterization of the Proliferating Cell Nuclear Antigen of Leishmania donovani Clinical Isolates and Its Association with Antimony Resistance"],"prefix":"10.1128","volume":"58","author":[{"given":"Rati","family":"Tandon","sequence":"first","affiliation":[{"name":"Division of Parasitology, Central Drug Research Institute, Lucknow, 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