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The Acidic Protein Binding Site Is Partially Hidden in the Free Saccharomyces cereVisiae Ribosomal Stalk Protein P0

dc.contributor.authorPérez-Fernández, Jorge
dc.contributor.authorRemacha, Miguel
dc.contributor.authorGarcía-Ballesta, Juan Pedro
dc.date.accessioned2025-01-07T07:20:37Z
dc.date.available2025-01-07T07:20:37Z
dc.date.issued2005-03-16
dc.description.abstractThe ribosomal stalk is essential for translation; however, its overall structure is poorly understood. Characterization of the region involved in the interactions between protein P0 and the 12kDa acidic proteins P1 and P2 is fundamental to understand the assembly and function of this structure in the eukaryotic ribosome. The acidic protein content is important for the ribosome efficiency and affects the translation of specific mRNAs. By usage of a series of progressively truncated fragments of protein P0 in the two-hybrid test, a region between positions 213 and 250 was identified as the minimal protein part able to interact with the acidic proteins. Extensions at either end affect the binding capacity of the fragment either positively or negatively depending on the number of added amino acids. Deletions inside the binding region confirm its in vivo relevance since they drastically reduce the P0 interacting capacity with the 12 kDa acidic proteins, which are severely reduced in the ribosome when the truncated protein is expressed in the cell. Moreover, recombinant His-tagged P0 fragments containing the binding site and bound to Ni2+-NTA columns can form a complex with the P1 and P2 proteins, which is able to bind elongation factor EF2. The results indicate the existence of a region in P0 that specifically interacts with the acidic proteins. These interactions are, however, hindered by the presence of neighbor protein domains, suggesting the need for conformational changes in the complete P0 to allow the assembly of the ribosomal stalk.es_ES
dc.description.sponsorshipMinisterio de Ciencia y Tecnología (Spain): BMC2003-03387 and GEN2001-4707-C08-05. European Commission Grant QLRT-2001-00892. Institutional grant from the Fundación Ramón Areces (Madrid).es_ES
dc.identifier.citationPérez-Fernández, J., Remacha, M. & Ballesta, J. P. The Acidic Protein Binding Site Is Partially Hidden in the Free Saccharomyces cerevisiae Ribosomal Stalk Protein P0. Biochemistry 44, 5532–5540 (2005).es_ES
dc.identifier.issn0006-2960es_ES
dc.identifier.other10.1021/bi047332res_ES
dc.identifier.urihttps://pubs.acs.org/doi/10.1021/bi047332res_ES
dc.identifier.urihttps://hdl.handle.net/10953/3728
dc.language.isoenges_ES
dc.publisherAmerican Chemical Societyes_ES
dc.relation.ispartofBiochemistry [2005]; [44 ]:[5532–5540]es_ES
dc.rights.accessRightsinfo:eu-repo/semantics/openAccesses_ES
dc.subjectRibosomees_ES
dc.subjectRibosomal stalkes_ES
dc.subjectPhosphoproteinses_ES
dc.subjectTranslationes_ES
dc.subjectSaccharomyces cerevisiaees_ES
dc.subjectIn vitro reconstitutiones_ES
dc.subjectAntibioticses_ES
dc.titleThe Acidic Protein Binding Site Is Partially Hidden in the Free Saccharomyces cereVisiae Ribosomal Stalk Protein P0es_ES
dc.typeinfo:eu-repo/semantics/articlees_ES
dc.type.versioninfo:eu-repo/semantics/acceptedVersiones_ES

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