Biochemical and structural characterization of the RT domain of Leishmania sp. telomerase reverse transcriptase

Autor(es)

da Silva, Vitor Luiz ; de Paiva, Stephany Cacete ; de Oliveira, Hamine Cristina ; Fernandes, Carlos Alexandre H. ; Salvador, Guilherme Henrique Marchi ; Fontes, Marcos Roberto de M. ; Cano, Maria Isabel N.

• Tipo de documento: Artigo Científico
• Fonte: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS; v. 1867, n. 11, p. 10-pg., 2023-09-28.

Resumo

Background: The Leishmania genus comprises parasites that cause leishmaniasis, a neglected disease spread worldwide. Leishmania sp. telomeres are composed of TTAGGG repeats maintained by telomerase. In most eukaryotes, the enzyme minimal complex contains the TER (telomerase RNA) and the TERT (telomerase reverse transcriptase) components. The TERT holds the enzyme catalytic core and is formed by four structural and functional domains (TEN, Telomerase Essential N-terminal; TRBD, Telomerase RNA Binding Domain; RT, the reverse transcriptase domain and CTE, C-Terminal Extension domain). Methods and results: Amino acid sequence alignments, protein structure prediction analysis, and protein: nucleic acid interaction assays were used to show that the Leishmania major RT domain preserves the canonical structural elements found in higher eukaryotes, including the canonical motifs and the aspartic acid residues that stabilize the Mg2+ ion cofactor. Furthermore, amino acid substitutions specific to the Leishmania genus and partial conservation of the residues involved with nucleic acid interactions are shown. The purified recombinant Leishmania RT protein is biochemically active and interacts with the G-rich telomeric strand and the TER template sequence. Conclusion: Our results highlight that the telomerase catalysis mechanism is conserved in a pathogen of medical importance despite the structural peculiarities present in the parasite's RT domain. (AU)

• Processo FAPESP: 19/05958-4 - EMU concedido no processo 2013/24705-3: sistema de cromatografia líquido de alta pressão
• Beneficiário: Marcos Roberto de Mattos Fontes
• Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
• Processo FAPESP: 21/14798-0 - Estudos funcionais e estruturais da proteína ortóloga a PinX1 e sua influência nos telômeros de Leishmania spp
• Beneficiário: Stephany Cacete de Paiva
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• 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: 19/05239-8 - Estudo comparativo entre diferentes técnicas biofísicas visando aprofundar o entendimento do mecanismo de importação nuclear
• Beneficiário: Hamine Cristina de Oliveira
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto