A short ncRNA modulates gene expression and affects stress response and parasite differentiation in Leishmania braziliensis
The protozoan parasite Leishmania spp. is a causative agent of leishmaniasis, a disease that affects millions of people in more than 80 countries worldwide. Apart from its medical relevance, this organism has a genetic organization that is unique among eukaryotes. Studies of the mechanisms regulatin...
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Frontiers Media S.A.
2025-02-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| author | José C. Quilles Caroline R. Espada Lissur A. Orsine Tânia A. Defina Letícia Almeida Fabíola Holetz Angela K. Cruz |
| author_facet | José C. Quilles Caroline R. Espada Lissur A. Orsine Tânia A. Defina Letícia Almeida Fabíola Holetz Angela K. Cruz |
| author_sort | José C. Quilles |
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| description | The protozoan parasite Leishmania spp. is a causative agent of leishmaniasis, a disease that affects millions of people in more than 80 countries worldwide. Apart from its medical relevance, this organism has a genetic organization that is unique among eukaryotes. Studies of the mechanisms regulating gene expression in Leishmania led us to investigate noncoding RNAs (ncRNAs) as regulatory elements. We previously identified differentially expressed (DE) ncRNAs in Leishmania braziliensis with potential roles in the parasite biology and development. Herein, we present a functional analysis of one such DE ncRNA, the 147-nucleotide-long transcript ncRNA97, which is preferentially expressed in amastigotes, the replicative form within mammalian phagocytes. By RT-qPCR the ncRNA97 was detected in greater quantities in the nucleus under physiological conditions and in the cytoplasm under nutritional stress. Interestingly, the transcript is protected at the 5’ end but is not processed by the canonical trypanosomatid trans-splicing mechanism, according to the RNA circularization assay. ncRNA97 knockout (KO) and addback (AB) transfectants were generated and subjected to phenotypic analysis, which revealed that the lack of ncRNA97 impairs the starvation response and differentiation to the infective form. Comparative transcriptomics of ncRNA97KO and parental cells revealed that transcripts encoding amastigote-specific proteins were affected. This pioneering work demonstrates that ncRNAs contribute to the developmental regulatory mechanisms of Leishmania. |
| format | Article |
| id | doaj-art-e0c7255cc79643f0aa906eb92c10f19e |
| institution | Kabale University |
| issn | 2235-2988 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj-art-e0c7255cc79643f0aa906eb92c10f19e2025-02-05T07:32:32ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-02-011510.3389/fcimb.2025.15139081513908A short ncRNA modulates gene expression and affects stress response and parasite differentiation in Leishmania braziliensisJosé C. Quilles0Caroline R. Espada1Lissur A. Orsine2Tânia A. Defina3Letícia Almeida4Fabíola Holetz5Angela K. Cruz6Laboratory de Molecular Parasitology, Department of Cell and Molecular Biology, Ribeirão Preto Medical School, FMRP/USP – University of São Paulo, Ribeirão Preto, SP, BrazilLaboratory de Molecular Parasitology, Department of Cell and Molecular Biology, Ribeirão Preto Medical School, FMRP/USP – University of São Paulo, Ribeirão Preto, SP, BrazilLaboratory de Molecular Parasitology, Department of Cell and Molecular Biology, Ribeirão Preto Medical School, FMRP/USP – University of São Paulo, Ribeirão Preto, SP, BrazilLaboratory de Molecular Parasitology, Department of Cell and Molecular Biology, Ribeirão Preto Medical School, FMRP/USP – University of São Paulo, Ribeirão Preto, SP, BrazilLaboratory de Molecular Parasitology, Department of Cell and Molecular Biology, Ribeirão Preto Medical School, FMRP/USP – University of São Paulo, Ribeirão Preto, SP, BrazilLaboratory of Gene Expression Regulation, Carlos Chagas Institute, Oswaldo Cruz Foundation, Curitiba, PR, BrazilLaboratory de Molecular Parasitology, Department of Cell and Molecular Biology, Ribeirão Preto Medical School, FMRP/USP – University of São Paulo, Ribeirão Preto, SP, BrazilThe protozoan parasite Leishmania spp. is a causative agent of leishmaniasis, a disease that affects millions of people in more than 80 countries worldwide. Apart from its medical relevance, this organism has a genetic organization that is unique among eukaryotes. Studies of the mechanisms regulating gene expression in Leishmania led us to investigate noncoding RNAs (ncRNAs) as regulatory elements. We previously identified differentially expressed (DE) ncRNAs in Leishmania braziliensis with potential roles in the parasite biology and development. Herein, we present a functional analysis of one such DE ncRNA, the 147-nucleotide-long transcript ncRNA97, which is preferentially expressed in amastigotes, the replicative form within mammalian phagocytes. By RT-qPCR the ncRNA97 was detected in greater quantities in the nucleus under physiological conditions and in the cytoplasm under nutritional stress. Interestingly, the transcript is protected at the 5’ end but is not processed by the canonical trypanosomatid trans-splicing mechanism, according to the RNA circularization assay. ncRNA97 knockout (KO) and addback (AB) transfectants were generated and subjected to phenotypic analysis, which revealed that the lack of ncRNA97 impairs the starvation response and differentiation to the infective form. Comparative transcriptomics of ncRNA97KO and parental cells revealed that transcripts encoding amastigote-specific proteins were affected. This pioneering work demonstrates that ncRNAs contribute to the developmental regulatory mechanisms of Leishmania.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1513908/fullnoncoding RNAleishmaniagene expressionmetacyclogenesisnutritional stress |
| spellingShingle | José C. Quilles Caroline R. Espada Lissur A. Orsine Tânia A. Defina Letícia Almeida Fabíola Holetz Angela K. Cruz A short ncRNA modulates gene expression and affects stress response and parasite differentiation in Leishmania braziliensis Frontiers in Cellular and Infection Microbiology noncoding RNA leishmania gene expression metacyclogenesis nutritional stress |
| title | A short ncRNA modulates gene expression and affects stress response and parasite differentiation in Leishmania braziliensis |
| title_full | A short ncRNA modulates gene expression and affects stress response and parasite differentiation in Leishmania braziliensis |
| title_fullStr | A short ncRNA modulates gene expression and affects stress response and parasite differentiation in Leishmania braziliensis |
| title_full_unstemmed | A short ncRNA modulates gene expression and affects stress response and parasite differentiation in Leishmania braziliensis |
| title_short | A short ncRNA modulates gene expression and affects stress response and parasite differentiation in Leishmania braziliensis |
| title_sort | short ncrna modulates gene expression and affects stress response and parasite differentiation in leishmania braziliensis |
| topic | noncoding RNA leishmania gene expression metacyclogenesis nutritional stress |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2025.1513908/full |
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