A single N1-methyladenosine on the large ribosomal subunit rRNA impacts locally its structure and the translation of key metabolic enzymes

Abstract The entire chemical modification repertoire of yeast ribosomal RNAs and the enzymes responsible for it have recently been identified. Nonetheless, in most cases the precise roles played by these chemical modifications in ribosome structure, function and regulation remain totally unclear. Pr...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sunny Sharma, Johannes David Hartmann, Peter Watzinger, Arvid Klepper, Christian Peifer, Peter Kötter, Denis L. J. Lafontaine, Karl-Dieter Entian
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/85152bebff554c44bd499cceadd25d67
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:85152bebff554c44bd499cceadd25d67
record_format	dspace
spelling	oai:doaj.org-article:85152bebff554c44bd499cceadd25d672021-12-02T15:08:26ZA single N1-methyladenosine on the large ribosomal subunit rRNA impacts locally its structure and the translation of key metabolic enzymes10.1038/s41598-018-30383-z2045-2322https://doaj.org/article/85152bebff554c44bd499cceadd25d672018-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-30383-zhttps://doaj.org/toc/2045-2322Abstract The entire chemical modification repertoire of yeast ribosomal RNAs and the enzymes responsible for it have recently been identified. Nonetheless, in most cases the precise roles played by these chemical modifications in ribosome structure, function and regulation remain totally unclear. Previously, we demonstrated that yeast Rrp8 methylates m1A645 of 25S rRNA in yeast. Here, using mung bean nuclease protection assays in combination with quantitative RP-HPLC and primer extension, we report that 25S/28S rRNA of S. pombe, C. albicans and humans also contain a single m1A methylation in the helix 25.1. We characterized nucleomethylin (NML) as a human homolog of yeast Rrp8 and demonstrate that NML catalyzes the m1A1322 methylation of 28S rRNA in humans. Our in vivo structural probing of 25S rRNA, using both DMS and SHAPE, revealed that the loss of the Rrp8-catalyzed m1A modification alters the conformation of domain I of yeast 25S rRNA causing translation initiation defects detectable as halfmers formation, likely because of incompetent loading of 60S on the 43S-preinitiation complex. Quantitative proteomic analysis of the yeast Δrrp8 mutant strain using 2D-DIGE, revealed that loss of m1A645 impacts production of specific set of proteins involved in carbohydrate metabolism, translation and ribosome synthesis. In mouse, NML has been characterized as a metabolic disease-associated gene linked to obesity. Our findings in yeast also point to a role of Rrp8 in primary metabolism. In conclusion, the m1A modification is crucial for maintaining an optimal 60S conformation, which in turn is important for regulating the production of key metabolic enzymes.Sunny SharmaJohannes David HartmannPeter WatzingerArvid KlepperChristian PeiferPeter KötterDenis L. J. LafontaineKarl-Dieter EntianNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-16 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sunny Sharma Johannes David Hartmann Peter Watzinger Arvid Klepper Christian Peifer Peter Kötter Denis L. J. Lafontaine Karl-Dieter Entian A single N1-methyladenosine on the large ribosomal subunit rRNA impacts locally its structure and the translation of key metabolic enzymes
description	Abstract The entire chemical modification repertoire of yeast ribosomal RNAs and the enzymes responsible for it have recently been identified. Nonetheless, in most cases the precise roles played by these chemical modifications in ribosome structure, function and regulation remain totally unclear. Previously, we demonstrated that yeast Rrp8 methylates m1A645 of 25S rRNA in yeast. Here, using mung bean nuclease protection assays in combination with quantitative RP-HPLC and primer extension, we report that 25S/28S rRNA of S. pombe, C. albicans and humans also contain a single m1A methylation in the helix 25.1. We characterized nucleomethylin (NML) as a human homolog of yeast Rrp8 and demonstrate that NML catalyzes the m1A1322 methylation of 28S rRNA in humans. Our in vivo structural probing of 25S rRNA, using both DMS and SHAPE, revealed that the loss of the Rrp8-catalyzed m1A modification alters the conformation of domain I of yeast 25S rRNA causing translation initiation defects detectable as halfmers formation, likely because of incompetent loading of 60S on the 43S-preinitiation complex. Quantitative proteomic analysis of the yeast Δrrp8 mutant strain using 2D-DIGE, revealed that loss of m1A645 impacts production of specific set of proteins involved in carbohydrate metabolism, translation and ribosome synthesis. In mouse, NML has been characterized as a metabolic disease-associated gene linked to obesity. Our findings in yeast also point to a role of Rrp8 in primary metabolism. In conclusion, the m1A modification is crucial for maintaining an optimal 60S conformation, which in turn is important for regulating the production of key metabolic enzymes.
format	article
author	Sunny Sharma Johannes David Hartmann Peter Watzinger Arvid Klepper Christian Peifer Peter Kötter Denis L. J. Lafontaine Karl-Dieter Entian
author_facet	Sunny Sharma Johannes David Hartmann Peter Watzinger Arvid Klepper Christian Peifer Peter Kötter Denis L. J. Lafontaine Karl-Dieter Entian
author_sort	Sunny Sharma
title	A single N1-methyladenosine on the large ribosomal subunit rRNA impacts locally its structure and the translation of key metabolic enzymes
title_short	A single N1-methyladenosine on the large ribosomal subunit rRNA impacts locally its structure and the translation of key metabolic enzymes
title_full	A single N1-methyladenosine on the large ribosomal subunit rRNA impacts locally its structure and the translation of key metabolic enzymes
title_fullStr	A single N1-methyladenosine on the large ribosomal subunit rRNA impacts locally its structure and the translation of key metabolic enzymes
title_full_unstemmed	A single N1-methyladenosine on the large ribosomal subunit rRNA impacts locally its structure and the translation of key metabolic enzymes
title_sort	single n1-methyladenosine on the large ribosomal subunit rrna impacts locally its structure and the translation of key metabolic enzymes
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/85152bebff554c44bd499cceadd25d67
work_keys_str_mv	AT sunnysharma asinglen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT johannesdavidhartmann asinglen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT peterwatzinger asinglen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT arvidklepper asinglen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT christianpeifer asinglen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT peterkotter asinglen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT denisljlafontaine asinglen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT karldieterentian asinglen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT sunnysharma singlen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT johannesdavidhartmann singlen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT peterwatzinger singlen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT arvidklepper singlen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT christianpeifer singlen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT peterkotter singlen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT denisljlafontaine singlen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes AT karldieterentian singlen1methyladenosineonthelargeribosomalsubunitrrnaimpactslocallyitsstructureandthetranslationofkeymetabolicenzymes
_version_	1718388131221209088

A single N1-methyladenosine on the large ribosomal subunit rRNA impacts locally its structure and the translation of key metabolic enzymes

Ejemplares similares