Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae

Ribosome biogenesis requires > 100 nonribosomal proteins, which are associated with different preribosomal particles. The substrates, the interacting partners, and the timing of action of most of these proteins are largely unknown. To elucidate the functional environment of the putative ATP-depen...

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Autores: Valle Rosado, Iván, Cruz Díaz, Jesús de la
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2004
País:España
Institución:Universidad de Sevilla (US)
Repositorio:idUS. Depósito de Investigación de la Universidad de Sevilla
OAI Identifier:oai:idus.us.es:11441/143526
Acceso en línea:https://hdl.handle.net/11441/143526
https://doi.org/10.1261/rna.7340404
Access Level:acceso abierto
Palabra clave:DEAD-box RNA helicase
Nucleolus
Preribosomal particles
Ribosome biogenesis
Yeast genetics
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spelling Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiaeValle Rosado, IvánCruz Díaz, Jesús de laDEAD-box RNA helicaseNucleolusPreribosomal particlesRibosome biogenesisYeast geneticsRibosome biogenesis requires > 100 nonribosomal proteins, which are associated with different preribosomal particles. The substrates, the interacting partners, and the timing of action of most of these proteins are largely unknown. To elucidate the functional environment of the putative ATP-dependent RNA helicase Dbp6p from Saccharomyces cerevisiae, which is required for 60S ribosomal subunit assembly, we have previously performed a synthetic lethal screen and thereby revealed a genetic interaction network between Dbp6p, Rp13p, Nop8p, and the novel Rsa3p. In this report, we extended the characterization of this functional network by performing a synthetic lethal screen with the rsa3 null allele. This screen identified the so far uncharacterized Npa1p (YKL014C). Polysome profile analysis indicates that there is a deficit of 60S ribosomal subunits and an accumulation of half-mer polysomes in the slowly growing npa1-1 mutant. Northern blotting and primer extension analysis shows that the npa1-1 mutation negatively affects processing of all 27S pre-rRNAs and the normal accumulation of both mature 25S and 5.8S rRNAs. In addition, 27SA2 pre-rRNA is prematurely cleaved at site C 2. Moreover, GFP-tagged Npa1p localizes predominantly to the nucleolus and sediments with large complexes in sucrose gradients, which most likely correspond to pre-60S ribosomal particles. We conclude that Npa1p is required for ribosome biogenesis and operates in the same functional environment of Rsa3p and Dbp6p during early maturation of 60S ribosomal subunits.Ministerio de Ciencia y Tecnología BMC2001-2660Junta de Andalucía CVI271Cold Spring Harbor Laboratory PressGenéticaMinisterio de Ciencia Y Tecnología (MCYT). EspañaJunta de Andalucía2004info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttps://hdl.handle.net/11441/143526https://doi.org/10.1261/rna.7340404reponame:idUS. Depósito de Investigación de la Universidad de Sevillainstname:Universidad de Sevilla (US)InglésRNA, 10 (7), 1073-1083.BMC2001-2660CVI271https://doi.org/10.1261/rna.7340404info:eu-repo/semantics/openAccessoai:idus.us.es:11441/1435262026-06-17T12:51:07Z
dc.title.none.fl_str_mv Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae
title Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae
spellingShingle Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae
Valle Rosado, Iván
DEAD-box RNA helicase
Nucleolus
Preribosomal particles
Ribosome biogenesis
Yeast genetics
title_short Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae
title_full Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae
title_fullStr Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae
title_full_unstemmed Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae
title_sort Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae
dc.creator.none.fl_str_mv Valle Rosado, Iván
Cruz Díaz, Jesús de la
author Valle Rosado, Iván
author_facet Valle Rosado, Iván
Cruz Díaz, Jesús de la
author_role author
author2 Cruz Díaz, Jesús de la
author2_role author
dc.contributor.none.fl_str_mv Genética
Ministerio de Ciencia Y Tecnología (MCYT). España
Junta de Andalucía
dc.subject.none.fl_str_mv DEAD-box RNA helicase
Nucleolus
Preribosomal particles
Ribosome biogenesis
Yeast genetics
topic DEAD-box RNA helicase
Nucleolus
Preribosomal particles
Ribosome biogenesis
Yeast genetics
description Ribosome biogenesis requires > 100 nonribosomal proteins, which are associated with different preribosomal particles. The substrates, the interacting partners, and the timing of action of most of these proteins are largely unknown. To elucidate the functional environment of the putative ATP-dependent RNA helicase Dbp6p from Saccharomyces cerevisiae, which is required for 60S ribosomal subunit assembly, we have previously performed a synthetic lethal screen and thereby revealed a genetic interaction network between Dbp6p, Rp13p, Nop8p, and the novel Rsa3p. In this report, we extended the characterization of this functional network by performing a synthetic lethal screen with the rsa3 null allele. This screen identified the so far uncharacterized Npa1p (YKL014C). Polysome profile analysis indicates that there is a deficit of 60S ribosomal subunits and an accumulation of half-mer polysomes in the slowly growing npa1-1 mutant. Northern blotting and primer extension analysis shows that the npa1-1 mutation negatively affects processing of all 27S pre-rRNAs and the normal accumulation of both mature 25S and 5.8S rRNAs. In addition, 27SA2 pre-rRNA is prematurely cleaved at site C 2. Moreover, GFP-tagged Npa1p localizes predominantly to the nucleolus and sediments with large complexes in sucrose gradients, which most likely correspond to pre-60S ribosomal particles. We conclude that Npa1p is required for ribosome biogenesis and operates in the same functional environment of Rsa3p and Dbp6p during early maturation of 60S ribosomal subunits.
publishDate 2004
dc.date.none.fl_str_mv 2004
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/11441/143526
https://doi.org/10.1261/rna.7340404
url https://hdl.handle.net/11441/143526
https://doi.org/10.1261/rna.7340404
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv RNA, 10 (7), 1073-1083.
BMC2001-2660
CVI271
https://doi.org/10.1261/rna.7340404
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
dc.source.none.fl_str_mv reponame:idUS. Depósito de Investigación de la Universidad de Sevilla
instname:Universidad de Sevilla (US)
instname_str Universidad de Sevilla (US)
reponame_str idUS. Depósito de Investigación de la Universidad de Sevilla
collection idUS. Depósito de Investigación de la Universidad de Sevilla
repository.name.fl_str_mv
repository.mail.fl_str_mv
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