A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis

Autor(es):Mostrar menos -

Fernandes, Carlos A. H. ^{[1, 2]} ; Morea, Edna Gicela O. ^[3] ; dos Santos, Gabriel A. ^[3] ; da Silva, Vitor L. ^[3] ; Vieira, Marina Roveri ^[3] ; Viviescas, Maria Alejandra ^[3] ; Chatain, Jean ^[4] ; Vadel, Aurelie ^[4] ; Saintome, Carole ^{[4, 5]} ; Fontes, Marcos Roberto M. ^[2] ; Cano, Maria Isabel Nogueira ^[3]

Afiliação do(s) autor(es)

^[1] Ecole Normale Super Paris Saclay, Lab Biol & Pharmacol Appl, Cachan - France

^[2] Sao Paulo State Univ UNESP, Biosci Inst, Dept Biophys & Pharmacol, Botucatu, SP - Brazil

^[3] Sao Paulo State Univ UNESP, Dept Chem & Biol Sci, Botucatu, SP - Brazil

^[4] INSERM U1154, MNHN CNRS UMR 7196, 43 Rue Cuvier, F-75005 Paris - France

^[5] Sorbonne Univ, UFR927, 4 Pl Jussieu, F-75005 Paris - France

• Tipo de documento: Artigo Científico
• Fonte: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS; v. 1864, n. 7 JUL 2020.
• Citações Web of Science: 0

Resumo

Background: Telomeres are chromosome end structures important in the maintenance of genome homeostasis. They are replenished by the action of telomerase and associated proteins, such as the OB (oligonucleotide/oligosaccharide-binding)-fold containing telomere-end binding proteins (TEBP) which plays an essential role in telomere maintenance and protection. The nature of TEBPs is well known in higher and some primitive eukaryotes, but it remains undetermined in trypanosomatids. Previous in silico searches have shown that there are no homologs of the classical TEPBs in trypanosomatids, including Leishmania sp. However, Replication Protein A subunit 1 (RPA-1), an OB-fold containing DNA-binding protein, was found co-localized with trypanosomatids telomeres and showed a high preference for the telomeric G-rich strand. Methods and results: We predicted the absence of structural homologs of OB-fold containing TEBPs in the Leishmania sp. genome using structural comparisons. We demonstrated by molecular docking that the ssDNA binding mode of LaRPA-1 shares features with the higher eukaryotes POT1 and RPA-1 crystal structures ssDNA binding mode. Using fluorescence spectroscopy, protein-DNA interaction assays, and FRET, we respectively show that LaRPA-1 shares some telomeric functions with the classical TEBPs since it can bind at least one telomeric repeat, protect the telomeric G-rich DNA from 3'-5' Exonuclease I digestion, and unfold telomeric G-quadruplex. Conclusions: Our results suggest that RPA-1 emerges as a TEBP in trypanosomatids, and in this context, we present two possible evolutionary landscapes of trypanosomatids RPA-1 that could reflect upon the evolution of OB-fold containing TEBPs from all eukaryotes. (AU)

• Processo FAPESP: 15/17286-0 - Estudos estruturais e funcionais objetivando compreender o papel de toxinas de venenos de serpentes e de como inibir a sua atividade biológica
• Beneficiário: Marcos Roberto de Mattos Fontes
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• 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: 19/11496-3 - 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