Insights into the role of endonuclease V in RNA metabolism in Trypanosoma brucei

Abstract Inosine may arise in DNA as a result of oxidative deamination of adenine or misincorporation of deoxyinosine triphosphate during replication. On the other hand, the occurrence of inosine in RNA is considered a normal and essential modification induced by specific adenosine deaminases acting...

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Autores principales:	Daniel García-Caballero, Guiomar Pérez-Moreno, Antonio M. Estévez, Luis Miguel Ruíz-Pérez, Antonio E. Vidal, Dolores González-Pacanowska
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e1476135eb1242f79c3d5f4e76ea151d
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spelling	oai:doaj.org-article:e1476135eb1242f79c3d5f4e76ea151d2021-12-02T12:32:43ZInsights into the role of endonuclease V in RNA metabolism in Trypanosoma brucei10.1038/s41598-017-08910-12045-2322https://doaj.org/article/e1476135eb1242f79c3d5f4e76ea151d2017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08910-1https://doaj.org/toc/2045-2322Abstract Inosine may arise in DNA as a result of oxidative deamination of adenine or misincorporation of deoxyinosine triphosphate during replication. On the other hand, the occurrence of inosine in RNA is considered a normal and essential modification induced by specific adenosine deaminases acting on mRNA and tRNA. In prokaryotes, endonuclease V (EndoV) can recognize and cleave inosine-containing DNA. In contrast, mammalian EndoVs preferentially cleave inosine-containing RNA, suggesting a role in RNA metabolism for the eukaryotic members of this protein family. We have performed a biochemical characterization of EndoV from the protozoan parasite Trypanosoma brucei. In vitro, TbEndoV efficiently processes single-stranded RNA oligonucleotides with inosine, including A to I-edited tRNA-like substrates but exhibits weak activity over DNA, except when a ribonucleotide is placed 3′ to the inosine. Immunolocalization studies performed in procyclic forms indicate that TbEndoV is mainly cytosolic yet upon nutritional stress it redistributes and accumulates in stress granules colocalizing with the DEAD-box helicase TbDhh1. RNAi-mediated depletion of TbEndoV results in moderate growth defects in procyclic cells while the two EndoV alleles could be readily knocked out in bloodstream forms. Taken together, these observations suggest an important role of TbEndoV in RNA metabolism in procyclic forms of the parasite.Daniel García-CaballeroGuiomar Pérez-MorenoAntonio M. EstévezLuis Miguel Ruíz-PérezAntonio E. VidalDolores González-PacanowskaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-15 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Daniel García-Caballero Guiomar Pérez-Moreno Antonio M. Estévez Luis Miguel Ruíz-Pérez Antonio E. Vidal Dolores González-Pacanowska Insights into the role of endonuclease V in RNA metabolism in Trypanosoma brucei
description	Abstract Inosine may arise in DNA as a result of oxidative deamination of adenine or misincorporation of deoxyinosine triphosphate during replication. On the other hand, the occurrence of inosine in RNA is considered a normal and essential modification induced by specific adenosine deaminases acting on mRNA and tRNA. In prokaryotes, endonuclease V (EndoV) can recognize and cleave inosine-containing DNA. In contrast, mammalian EndoVs preferentially cleave inosine-containing RNA, suggesting a role in RNA metabolism for the eukaryotic members of this protein family. We have performed a biochemical characterization of EndoV from the protozoan parasite Trypanosoma brucei. In vitro, TbEndoV efficiently processes single-stranded RNA oligonucleotides with inosine, including A to I-edited tRNA-like substrates but exhibits weak activity over DNA, except when a ribonucleotide is placed 3′ to the inosine. Immunolocalization studies performed in procyclic forms indicate that TbEndoV is mainly cytosolic yet upon nutritional stress it redistributes and accumulates in stress granules colocalizing with the DEAD-box helicase TbDhh1. RNAi-mediated depletion of TbEndoV results in moderate growth defects in procyclic cells while the two EndoV alleles could be readily knocked out in bloodstream forms. Taken together, these observations suggest an important role of TbEndoV in RNA metabolism in procyclic forms of the parasite.
format	article
author	Daniel García-Caballero Guiomar Pérez-Moreno Antonio M. Estévez Luis Miguel Ruíz-Pérez Antonio E. Vidal Dolores González-Pacanowska
author_facet	Daniel García-Caballero Guiomar Pérez-Moreno Antonio M. Estévez Luis Miguel Ruíz-Pérez Antonio E. Vidal Dolores González-Pacanowska
author_sort	Daniel García-Caballero
title	Insights into the role of endonuclease V in RNA metabolism in Trypanosoma brucei
title_short	Insights into the role of endonuclease V in RNA metabolism in Trypanosoma brucei
title_full	Insights into the role of endonuclease V in RNA metabolism in Trypanosoma brucei
title_fullStr	Insights into the role of endonuclease V in RNA metabolism in Trypanosoma brucei
title_full_unstemmed	Insights into the role of endonuclease V in RNA metabolism in Trypanosoma brucei
title_sort	insights into the role of endonuclease v in rna metabolism in trypanosoma brucei
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/e1476135eb1242f79c3d5f4e76ea151d
work_keys_str_mv	AT danielgarciacaballero insightsintotheroleofendonucleasevinrnametabolismintrypanosomabrucei AT guiomarperezmoreno insightsintotheroleofendonucleasevinrnametabolismintrypanosomabrucei AT antoniomestevez insightsintotheroleofendonucleasevinrnametabolismintrypanosomabrucei AT luismiguelruizperez insightsintotheroleofendonucleasevinrnametabolismintrypanosomabrucei AT antonioevidal insightsintotheroleofendonucleasevinrnametabolismintrypanosomabrucei AT doloresgonzalezpacanowska insightsintotheroleofendonucleasevinrnametabolismintrypanosomabrucei
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Insights into the role of endonuclease V in RNA metabolism in Trypanosoma brucei

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