RNase III Domain of KREPB9 and KREPB10 Association with Editosomes in <named-content content-type="genus-species">Trypanosoma brucei</named-content>

ABSTRACT Editosomes are the multiprotein complexes that catalyze the insertion and deletion of uridines to create translatable mRNAs in the mitochondria of kinetoplastids. Recognition and cleavage of a broad diversity of RNA substrates in vivo require three functionally distinct RNase III-type endon...

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Autores principales:	Jason Carnes, Suzanne M. McDermott, Kenneth Stuart
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	RNA editing RNase III Trypanosoma brucei editosome Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ee24059b884847dbb972ed5ed619f950
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spelling	oai:doaj.org-article:ee24059b884847dbb972ed5ed619f9502021-11-15T15:22:02ZRNase III Domain of KREPB9 and KREPB10 Association with Editosomes in <named-content content-type="genus-species">Trypanosoma brucei</named-content>10.1128/mSphereDirect.00585-172379-5042https://doaj.org/article/ee24059b884847dbb972ed5ed619f9502018-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphereDirect.00585-17https://doaj.org/toc/2379-5042ABSTRACT Editosomes are the multiprotein complexes that catalyze the insertion and deletion of uridines to create translatable mRNAs in the mitochondria of kinetoplastids. Recognition and cleavage of a broad diversity of RNA substrates in vivo require three functionally distinct RNase III-type endonucleases, as well as five additional editosome proteins that contain noncatalytic RNase III domains. RNase III domains have recently been identified in the editosome accessory proteins KREPB9 and KREPB10, suggesting a role related to editing endonuclease function. In this report, we definitively show that KREPB9 and KREPB10 are not essential in either bloodstream-form parasites (BF) or procyclic-form parasites (PF) by creating null or conditional null cell lines. While preedited and edited transcripts are largely unaffected by the loss of KREPB9 in both PF and BF, loss of KREPB10 produces distinct responses in BF and PF. BF cells lacking KREPB10 also lack edited CYb, while PF cells have increased edited A6, RPS12, ND3, and COII after loss of KREPB10. We also demonstrate that mutation of the RNase III domain of either KREPB9 or KREPB10 results in decreased association with ~20S editosomes. Editosome interactions with KREPB9 and KREPB10 are therefore mediated by the noncatalytic RNase III domain, consistent with a role in endonuclease specialization in Trypanosoma brucei. IMPORTANCE Trypanosoma brucei is a protozoan parasite that causes African sleeping sickness. U insertion/deletion RNA editing in T. brucei generates mature mitochondrial mRNAs. Editing is essential for survival in mammalian hosts and tsetse fly vectors and is differentially regulated during the parasite life cycle. Three multiprotein “editosomes,” typified by exclusive RNase III endonucleases that act at distinct sites, catalyze editing. Here, we show that editosome accessory proteins KREPB9 and KREPB10 are not essential for mammalian blood- or insect-form parasite survival but have specific and differential effects on edited RNA abundance in different stages. We also characterize KREPB9 and KREPB10 noncatalytic RNase III domains and show they are essential for editosome association, potentially via dimerization with RNase III domains in other editosome proteins. This work enhances the understanding of distinct editosome and accessory protein functions, and thus differential editing, during the parasite life cycle and highlights the importance of RNase III domain interactions to editosome architecture.Jason CarnesSuzanne M. McDermottKenneth StuartAmerican Society for MicrobiologyarticleRNA editingRNase IIITrypanosoma bruceieditosomeMicrobiologyQR1-502ENmSphere, Vol 3, Iss 1 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	RNA editing RNase III Trypanosoma brucei editosome Microbiology QR1-502
spellingShingle	RNA editing RNase III Trypanosoma brucei editosome Microbiology QR1-502 Jason Carnes Suzanne M. McDermott Kenneth Stuart RNase III Domain of KREPB9 and KREPB10 Association with Editosomes in <named-content content-type="genus-species">Trypanosoma brucei</named-content>
description	ABSTRACT Editosomes are the multiprotein complexes that catalyze the insertion and deletion of uridines to create translatable mRNAs in the mitochondria of kinetoplastids. Recognition and cleavage of a broad diversity of RNA substrates in vivo require three functionally distinct RNase III-type endonucleases, as well as five additional editosome proteins that contain noncatalytic RNase III domains. RNase III domains have recently been identified in the editosome accessory proteins KREPB9 and KREPB10, suggesting a role related to editing endonuclease function. In this report, we definitively show that KREPB9 and KREPB10 are not essential in either bloodstream-form parasites (BF) or procyclic-form parasites (PF) by creating null or conditional null cell lines. While preedited and edited transcripts are largely unaffected by the loss of KREPB9 in both PF and BF, loss of KREPB10 produces distinct responses in BF and PF. BF cells lacking KREPB10 also lack edited CYb, while PF cells have increased edited A6, RPS12, ND3, and COII after loss of KREPB10. We also demonstrate that mutation of the RNase III domain of either KREPB9 or KREPB10 results in decreased association with ~20S editosomes. Editosome interactions with KREPB9 and KREPB10 are therefore mediated by the noncatalytic RNase III domain, consistent with a role in endonuclease specialization in Trypanosoma brucei. IMPORTANCE Trypanosoma brucei is a protozoan parasite that causes African sleeping sickness. U insertion/deletion RNA editing in T. brucei generates mature mitochondrial mRNAs. Editing is essential for survival in mammalian hosts and tsetse fly vectors and is differentially regulated during the parasite life cycle. Three multiprotein “editosomes,” typified by exclusive RNase III endonucleases that act at distinct sites, catalyze editing. Here, we show that editosome accessory proteins KREPB9 and KREPB10 are not essential for mammalian blood- or insect-form parasite survival but have specific and differential effects on edited RNA abundance in different stages. We also characterize KREPB9 and KREPB10 noncatalytic RNase III domains and show they are essential for editosome association, potentially via dimerization with RNase III domains in other editosome proteins. This work enhances the understanding of distinct editosome and accessory protein functions, and thus differential editing, during the parasite life cycle and highlights the importance of RNase III domain interactions to editosome architecture.
format	article
author	Jason Carnes Suzanne M. McDermott Kenneth Stuart
author_facet	Jason Carnes Suzanne M. McDermott Kenneth Stuart
author_sort	Jason Carnes
title	RNase III Domain of KREPB9 and KREPB10 Association with Editosomes in <named-content content-type="genus-species">Trypanosoma brucei</named-content>
title_short	RNase III Domain of KREPB9 and KREPB10 Association with Editosomes in <named-content content-type="genus-species">Trypanosoma brucei</named-content>
title_full	RNase III Domain of KREPB9 and KREPB10 Association with Editosomes in <named-content content-type="genus-species">Trypanosoma brucei</named-content>
title_fullStr	RNase III Domain of KREPB9 and KREPB10 Association with Editosomes in <named-content content-type="genus-species">Trypanosoma brucei</named-content>
title_full_unstemmed	RNase III Domain of KREPB9 and KREPB10 Association with Editosomes in <named-content content-type="genus-species">Trypanosoma brucei</named-content>
title_sort	rnase iii domain of krepb9 and krepb10 association with editosomes in <named-content content-type="genus-species">trypanosoma brucei</named-content>
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/ee24059b884847dbb972ed5ed619f950
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RNase III Domain of KREPB9 and KREPB10 Association with Editosomes in <named-content content-type="genus-species">Trypanosoma brucei</named-content>

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