A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.bbagen.2020.129607 http://hdl.handle.net/11449/200232 |
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. |
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A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensisG-quadruplexLeishmania sp.OB-foldRPA-1Telomere end-binding proteinBackground: 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.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Centre National de la Recherche ScientifiqueFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Institut National de la Santé et de la Recherche MédicaleMuséum National d'Histoire NaturelleCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Department of Biophysics and Pharmacology Biosciences Institute São Paulo State University (UNESP) – BotucatuLaboratoire de Biologie et Pharmacologie Appliquée École Normale Supérieure Paris-SaclayDepartment of Chemical and Biological Sciences São Paulo State University (UNESP) – BotucatuMNHN CNRS UMR 7196 INSERM U1154, 43 rue CuvierSorbonne Université UFR927, 4 place JussieuDepartment of Biophysics and Pharmacology Biosciences Institute São Paulo State University (UNESP) – BotucatuDepartment of Chemical and Biological Sciences São Paulo State University (UNESP) – BotucatuCAPES: 171895/2018-01FAPESP: 2015/17286-0FAPESP: 2015/18641-5FAPESP: 2018/0395-6FAPESP: 2018/04375-2FAPESP: 2019/11496-3CNPq: 305300/2006-7CAPES: 88881Universidade Estadual Paulista (Unesp)École Normale Supérieure Paris-SaclayINSERM U1154UFR927Fernandes, Carlos A.H. [UNESP]Morea, Edna Gicela O. [UNESP]dos Santos, Gabriel A. [UNESP]da Silva, Vitor L. [UNESP]Vieira, Marina Roveri [UNESP]Viviescas, Maria Alejandra [UNESP]Chatain, JeanVadel, AurélieSaintomé, CaroleFontes, Marcos Roberto M. [UNESP]Cano, Maria Isabel Nogueira [UNESP]2020-12-12T02:01:06Z2020-12-12T02:01:06Z2020-07-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1016/j.bbagen.2020.129607Biochimica et Biophysica Acta - General Subjects, v. 1864, n. 7, 2020.1872-80060304-4165http://hdl.handle.net/11449/20023210.1016/j.bbagen.2020.1296072-s2.0-85082711286Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBiochimica et Biophysica Acta - General Subjectsinfo:eu-repo/semantics/openAccess2021-10-23T12:31:47Zoai:repositorio.unesp.br:11449/200232Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462021-10-23T12:31:47Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis |
title |
A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis |
spellingShingle |
A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis Fernandes, Carlos A.H. [UNESP] G-quadruplex Leishmania sp. OB-fold RPA-1 Telomere end-binding protein |
title_short |
A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis |
title_full |
A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis |
title_fullStr |
A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis |
title_full_unstemmed |
A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis |
title_sort |
A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis |
author |
Fernandes, Carlos A.H. [UNESP] |
author_facet |
Fernandes, Carlos A.H. [UNESP] Morea, Edna Gicela O. [UNESP] dos Santos, Gabriel A. [UNESP] da Silva, Vitor L. [UNESP] Vieira, Marina Roveri [UNESP] Viviescas, Maria Alejandra [UNESP] Chatain, Jean Vadel, Aurélie Saintomé, Carole Fontes, Marcos Roberto M. [UNESP] Cano, Maria Isabel Nogueira [UNESP] |
author_role |
author |
author2 |
Morea, Edna Gicela O. [UNESP] dos Santos, Gabriel A. [UNESP] da Silva, Vitor L. [UNESP] Vieira, Marina Roveri [UNESP] Viviescas, Maria Alejandra [UNESP] Chatain, Jean Vadel, Aurélie Saintomé, Carole Fontes, Marcos Roberto M. [UNESP] Cano, Maria Isabel Nogueira [UNESP] |
author2_role |
author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) École Normale Supérieure Paris-Saclay INSERM U1154 UFR927 |
dc.contributor.author.fl_str_mv |
Fernandes, Carlos A.H. [UNESP] Morea, Edna Gicela O. [UNESP] dos Santos, Gabriel A. [UNESP] da Silva, Vitor L. [UNESP] Vieira, Marina Roveri [UNESP] Viviescas, Maria Alejandra [UNESP] Chatain, Jean Vadel, Aurélie Saintomé, Carole Fontes, Marcos Roberto M. [UNESP] Cano, Maria Isabel Nogueira [UNESP] |
dc.subject.por.fl_str_mv |
G-quadruplex Leishmania sp. OB-fold RPA-1 Telomere end-binding protein |
topic |
G-quadruplex Leishmania sp. OB-fold RPA-1 Telomere end-binding protein |
description |
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. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-12-12T02:01:06Z 2020-12-12T02:01:06Z 2020-07-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.bbagen.2020.129607 Biochimica et Biophysica Acta - General Subjects, v. 1864, n. 7, 2020. 1872-8006 0304-4165 http://hdl.handle.net/11449/200232 10.1016/j.bbagen.2020.129607 2-s2.0-85082711286 |
url |
http://dx.doi.org/10.1016/j.bbagen.2020.129607 http://hdl.handle.net/11449/200232 |
identifier_str_mv |
Biochimica et Biophysica Acta - General Subjects, v. 1864, n. 7, 2020. 1872-8006 0304-4165 10.1016/j.bbagen.2020.129607 2-s2.0-85082711286 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Biochimica et Biophysica Acta - General Subjects |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
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1797789241516752896 |