Deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating Amphotericin B resistance and survival of Leishmania donovani under oxidative stress
Leishmania donovani is the causative organism of the neglected human disease known as visceral leishmaniasis which is often fatal, if left untreated. The cysteine biosynthesis pathway of Leishmania may serve as a potential drug target because it is different from human host and regulates downstream...
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| Format: | Article |
| Language: | English |
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Elsevier
2017-08-01
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| Series: | Redox Biology |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231717301581 |
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| author | Kuljit Singh Vahab Ali Krishn Pratap Singh Parool Gupta Shashi S. Suman Ayan K. Ghosh Sanjiva Bimal Krishna Pandey Pradeep Das |
| author_facet | Kuljit Singh Vahab Ali Krishn Pratap Singh Parool Gupta Shashi S. Suman Ayan K. Ghosh Sanjiva Bimal Krishna Pandey Pradeep Das |
| author_sort | Kuljit Singh |
| collection | DOAJ |
| description | Leishmania donovani is the causative organism of the neglected human disease known as visceral leishmaniasis which is often fatal, if left untreated. The cysteine biosynthesis pathway of Leishmania may serve as a potential drug target because it is different from human host and regulates downstream components of redox metabolism of the parasites; essential for their survival, pathogenicity and drug resistance. However, despite the apparent dependency of redox metabolism of cysteine biosynthesis pathway, the role of L. donovani cysteine synthase (LdCS) in drug resistance and redox homeostasis has been unexplored. Herein, we report that over-expression of LdCS in Amphotericin B (Amp B) sensitive strain (S1-OE) modulates resistance towards oxidative stress and drug pressure. We observed that antioxidant enzyme activities were up-regulated in S1-OE parasites and these parasites alleviate intracellular reactive oxygen species (ROS) efficiently by maintaining the reduced thiol pool. In contrast to S1-OE parasites, Amp B sensitive strain (S1) showed higher levels of ROS which was positively correlated with the protein carbonylation levels and negatively correlated with cell viability. Moreover, further investigations showed that LdCS over-expression also augments the ROS-primed induction of LdCS-GFP as well as endogenous LdCS and thiol pathway proteins (LdTryS, LdTryR and LdcTXN) in L. donovani parasites; which probably aids in stress tolerance and drug resistance. In addition, the expression of LdCS was found to be up-regulated in Amp B resistant isolates and during infective stationary stages of growth and consistent with these observations, our ex vivo infectivity studies confirmed that LdCS over-expression enhances the infectivity of L. donovani parasites. Our results reveal a novel crosstalk between LdCS and thiol metabolic pathway proteins and demonstrate the crucial role of LdCS in drug resistance and redox homeostasis of Leishmania. Keywords: Leishmania, Cysteine synthase, Drug resistance, Thiol metabolism, Oxidative stress, Amphotericin B, Trypanothione |
| first_indexed | 2024-12-20T13:20:48Z |
| format | Article |
| id | doaj.art-41e1c0e7f7b342fa88520072dc84f9a6 |
| institution | Directory Open Access Journal |
| issn | 2213-2317 |
| language | English |
| last_indexed | 2024-12-20T13:20:48Z |
| publishDate | 2017-08-01 |
| publisher | Elsevier |
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| series | Redox Biology |
| spelling | doaj.art-41e1c0e7f7b342fa88520072dc84f9a62022-12-21T19:39:24ZengElsevierRedox Biology2213-23172017-08-0112350366Deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating Amphotericin B resistance and survival of Leishmania donovani under oxidative stressKuljit Singh0Vahab Ali1Krishn Pratap Singh2Parool Gupta3Shashi S. Suman4Ayan K. Ghosh5Sanjiva Bimal6Krishna Pandey7Pradeep Das8Laboratory of Molecular Biochemistry and Cell Biology, Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, EPIP Complex, Hajipur 844102, IndiaLaboratory of Molecular Biochemistry and Cell Biology, Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, EPIP Complex, Hajipur 844102, India; Corresponding author at: Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, India.Laboratory of Molecular Biochemistry and Cell Biology, Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, IndiaLaboratory of Molecular Biochemistry and Cell Biology, Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, IndiaLaboratory of Molecular Biochemistry and Cell Biology, Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, IndiaDepartment of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, IndiaDepartment of Immunology, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, IndiaDepartment of Clinical Medicine, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, IndiaDepartment of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, IndiaLeishmania donovani is the causative organism of the neglected human disease known as visceral leishmaniasis which is often fatal, if left untreated. The cysteine biosynthesis pathway of Leishmania may serve as a potential drug target because it is different from human host and regulates downstream components of redox metabolism of the parasites; essential for their survival, pathogenicity and drug resistance. However, despite the apparent dependency of redox metabolism of cysteine biosynthesis pathway, the role of L. donovani cysteine synthase (LdCS) in drug resistance and redox homeostasis has been unexplored. Herein, we report that over-expression of LdCS in Amphotericin B (Amp B) sensitive strain (S1-OE) modulates resistance towards oxidative stress and drug pressure. We observed that antioxidant enzyme activities were up-regulated in S1-OE parasites and these parasites alleviate intracellular reactive oxygen species (ROS) efficiently by maintaining the reduced thiol pool. In contrast to S1-OE parasites, Amp B sensitive strain (S1) showed higher levels of ROS which was positively correlated with the protein carbonylation levels and negatively correlated with cell viability. Moreover, further investigations showed that LdCS over-expression also augments the ROS-primed induction of LdCS-GFP as well as endogenous LdCS and thiol pathway proteins (LdTryS, LdTryR and LdcTXN) in L. donovani parasites; which probably aids in stress tolerance and drug resistance. In addition, the expression of LdCS was found to be up-regulated in Amp B resistant isolates and during infective stationary stages of growth and consistent with these observations, our ex vivo infectivity studies confirmed that LdCS over-expression enhances the infectivity of L. donovani parasites. Our results reveal a novel crosstalk between LdCS and thiol metabolic pathway proteins and demonstrate the crucial role of LdCS in drug resistance and redox homeostasis of Leishmania. Keywords: Leishmania, Cysteine synthase, Drug resistance, Thiol metabolism, Oxidative stress, Amphotericin B, Trypanothionehttp://www.sciencedirect.com/science/article/pii/S2213231717301581 |
| spellingShingle | Kuljit Singh Vahab Ali Krishn Pratap Singh Parool Gupta Shashi S. Suman Ayan K. Ghosh Sanjiva Bimal Krishna Pandey Pradeep Das Deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating Amphotericin B resistance and survival of Leishmania donovani under oxidative stress Redox Biology |
| title | Deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating Amphotericin B resistance and survival of Leishmania donovani under oxidative stress |
| title_full | Deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating Amphotericin B resistance and survival of Leishmania donovani under oxidative stress |
| title_fullStr | Deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating Amphotericin B resistance and survival of Leishmania donovani under oxidative stress |
| title_full_unstemmed | Deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating Amphotericin B resistance and survival of Leishmania donovani under oxidative stress |
| title_short | Deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating Amphotericin B resistance and survival of Leishmania donovani under oxidative stress |
| title_sort | deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating amphotericin b resistance and survival of leishmania donovani under oxidative stress |
| url | http://www.sciencedirect.com/science/article/pii/S2213231717301581 |
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