title: GTP-dependent formation of a ribonucleoprotein subcomplex required for ribosome biogenesis.
abstract: Ribosome biogenesis in eukaryotic organisms involves the coordinated assembly of 78 ribosomal proteins onto the four ribosomal RNAs, mediated by a host of trans-acting factors whose specific functions remain largely unknown. The essential GTPase Bms1, the putative endonuclease Rcl1 and the essential U3 small nucleolar RNA form a stable subcomplex thought to control an early step in the assembly of the 40S ribosomal subunit. Here, we provide a complete thermodynamic analysis of GTP-dependent subcomplex formation, revealing strong thermodynamic coupling of Rcl1, U3 small nucleolar RNA and GTP binding to Bms1 that is eliminated in the presence of GDP. The results suggest that Rcl1 activates Bms1 by promoting GDP/GTP exchange, analogous to ribosome-promoted nucleotide exchange within translation elongation factor EF-G. These and other data unveil thermodynamic similarities between Bms1 and the subgroup of GTPases involved in translation, providing evidence that parts of the ribosome assembly machinery may have evolved from the translation apparatus. This quantitative description of an early and essential step in pre-ribosome assembly provides a framework for elucidating the network of interactions between the Bms1 subcomplex and additional factors involved in ribosome biogenesis.
authors.affiliation: Department of Molecular and Cell Biology, University of California at Berkeley, 94720-3206, USA.
authors.forename: Katrin
authors.initials: K
authors.lastname: Karbstein
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Doudna
mesh_terms: D019204:GTP-Binding Proteins / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006165:Guanylyl Imidodiphosphate / Q000378:metabolism; D046911:Macromolecular Substances; D009687:Nuclear Proteins / Q000378:metabolism; D011485:Protein Binding; D020537:RNA, Small Nucleolar / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry* / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013816:Thermodynamics
chemical_list: C436684:Bms1 protein, S cerevisiae; D046911:Macromolecular Substances; D009687:Nuclear Proteins; C409342:RCL1 protein, S cerevisiae; D020537:RNA, Small Nucleolar; C117378:RNA, U3 small nucleolar; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D029701:Saccharomyces cerevisiae Proteins; D006153:Guanosine Diphosphate; D006165:Guanylyl Imidodiphosphate; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins