Exploring TERRA during Leishmania major developmental cycle and continuous in vitro passages

Autor(es)

Ortiz Morea, Edna Gicela ^[1] ; Vasconcelos, Elton Jose Rosas ^[2] ; Alves, Cristiane de Santis ^[1] ; Giorgio, Selma ^[3] ; Myler, Peter J. ^{[4, 5]} ; Langoni, Helio ^[6] ; Azzalin, Claus Maria ^[7] ; Nogueira Cano, Maria Isabel ^[1]

Afiliação do(s) autor(es)

^[1] Sao Paulo State Univ, Biosci Inst, Dept Chem & Biol Sci, UNESP, Botucatu, SP - Brazil

^[2] Univ Leeds, Leeds Omics, Leeds, W Yorkshire - England

^[3] Univ Estadual Campinas, Biol Inst, Dept Anim Biol, UNICAMP, Campinas, SP - Brazil

^[4] Univ Washington, Dept Global Hlth, Seattle, WA 98195 - USA

^[5] Univ Washington, Dept Biomed Informat & Med Educ, Seattle, WA 98195 - USA

^[6] Sao Paulo State Univ, UNESP, Vet Med Sch, Dept Publ Hlth, Botucatu, SP - Brazil

^[7] Inst Mol Med, Lisbon - Portugal

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 174, p. 573-586, MAR 31 2021.
• Citações Web of Science: 0

Resumo

Telomeres from different eukaryotes, including trypanosomatids, are transcribed into TERRA noncoding RNAs, crucial in regulating chromatin deposition and telomere length. TERRA is transcribed from the C-rich subtelomeric strand towards the 3'-ends of the telomeric array. Using bioinformatics, we confirmed the presence of subtelomeric splice acceptor sites at all L. major chromosome ends. Splice leader sequences positioned 5' upstream of L. major chromosomes subtelomeres were then mapped using SL-RNA-Seq libraries constructed from three independent parasite life stages and helped confirm TERRA expression from several chromosomes ends. Northern blots and RT-qPCR validated the results showing that L. major TERRA is processed by trans-splicing and polyadenylation coupled reactions. The number of transcripts varied with the parasite's life stage and continuous passages, being more abundant in the infective forms. However, no putative subtelomeric promoters involved in TERRA's transcriptional regulation were detected. In contrast, the observed changes in parasite's telomere length during development, suggest that differences in telomeric base J levels may control TERRA transcription in L. major. Also, TERRA-R loops detection, mainly in the infective forms, was suggestive of TERRA's involvement in telomere protection. Therefore, Leishmania TERRA shares conserved features with other eukaryotes and advances new telomere specific functions in a Public Health-impacting parasite. (C) 2021 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 19/11496-3 - PAPEL DO TERRA NA REGULAÇÃO DA MANUTENÇÃO DOS TELÔMEROS E NA SENESCÊNCIA REPLICATIVA EM Leishmania major
• Beneficiário: Edna Gicela Ortiz Morea
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 18/04375-2 - Estudos sobre a biogênese e composição do complexo ribonucleoprotéico da telomerase de Leishmania spp. e sua regulação
• Beneficiário: Maria Isabel Nogueira Cano
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 18/23302-6 - Leishmanioses de importância médica no Brasil: estudo de sistemas celulares e isolados clínicos de Leishmania infantum e L. braziliensis e avaliação de candidatos vacinais contra L. infantum
• Beneficiário: Selma Giorgio
• Modalidade de apoio: Auxílio à Pesquisa - Regular