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Computational discovery of conserved RNA structures and functional characterization of a structured lncRNA in Leishmania braziliensis

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Espadaa, Caroline R. ; Anthonb, Christian ; Magalhaesa, Rubens D. M. ; Junior, Jose Carlos Quilles ; Teles, Natalia M. M. ; Pais, Fabiano S. ; Orsine, Lissur A. ; de Almeida, Leticia ; Defina, Tania P. A. ; Dowle, Adam ; Gorodkin, Jan ; Walrad, Pegine B. ; Cruz, Angela K.
Total Authors: 13
Document type: Journal article
Source: NON-CODING RNA RESEARCH; v. 14, p. 14-pg., 2025-10-01.
Abstract

Leishmania parasites alternate between hosts, facing environmental changes that demand rapid gene expression adaptation. Lacking canonical RNA polymerase II promoters, transcription in these eukaryotes is polycistronic, with gene regulation occurring post-transcriptionally. Although non-coding RNAs (ncRNAs) have been identified in Leishmania transcriptomes, their functions remain unclear. Recognizing RNA structure's importance, we performed a genome-wide alignment of L. braziliensis and related species, identifying conserved RNA structures, 38 of which overlap with known ncRNAs. One such ncRNA, lncRNA45, was functionally characterized. Using a knockout cell line, we demonstrated that lncRNA45 is crucial for parasite fitness. Reintroducing the wild type lncRNA45 restored fitness, while a version with a single nucleotide substitution in the structured region did not. This mutation also altered RNA-protein interactions. These findings suggest that lncRNA45's regulatory role and protein interactions rely on its secondary structure. This study highlights the significance of structured lncRNAs in Leishmania biology and their potential as therapeutic targets. Further research into these ncRNAs could uncover new parasite regulation mechanisms and inspire novel treatment strategies. (AU)

FAPESP's process: 21/15182-3 - Monitoring processing, localisation and function of differentially expressed ncRNAs in Leishmania braziliensis by multi-colour RNA FISH on subcellular resolution
Grantee:José Carlos Quilles Junior
Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
FAPESP's process: 20/00087-2 - Functional validation of potential non-coding RNAs differentially expressed during Leishmania viannia braziliensis life cycle
Grantee:Caroline Ricce Espada
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 21/10043-5 - Comparative computational analyses of transcriptomes and large-scale identification of interactions between RNA-binding proteins and ncRNAs and repair/recombination proteins and DNA
Grantee:Lissur Azevedo Orsine
Support Opportunities: Scholarships in Brazil - Technical Training Program - Technical Training
FAPESP's process: 17/19040-3 - Use of high-content screening to evaluate drug leishmanicidal activity in macrophages
Grantee:Leticia de Almeida
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 15/13618-8 - Regulating the trans-regulators: investigating the PRMT7 molecular pathway as an epigenetic regulator of Leishmania virulence
Grantee:Angela Kaysel Cruz
Support Opportunities: Research Projects - Thematic Grants
FAPESP's process: 22/10270-4 - Investigation of non-coding RNA-protein interactions in Leishmania (Viannia) braziliensis: identifying ligands and dissecting binding profiles
Grantee:Caroline Ricce Espada
Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
FAPESP's process: 23/18057-0 - Evaluation of the role of parasite's Argonaute on infection by Leishmania braziliensis
Grantee:Lissur Azevedo Orsine
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 19/18607-5 - Computational identification and structural characterization of non-coding RNA in Leishmania braziliensis
Grantee:Rubens Daniel Miserani Magalhães
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 20/00088-9 - Epigenetic regulation of Leishmania gene expression
Grantee:José Carlos Quilles Junior
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 18/14398-0 - UK:Brazil Joint Centre Partnership in Leishmaniasis (JCPiL)
Grantee:Angela Kaysel Cruz
Support Opportunities: Research Projects - Thematic Grants