title: Analysis of the ribosome large subunit assembly and 23 S rRNA stability in vivo.
abstract: The ability of mutant 23 S ribosomal RNA to form particles with proteins of the large ribosomal subunit in vivo was studied. A series of overlapping deletions covering the entire 23 S rRNA, were constructed in the plasmid copy of an E. coli 23 S rRNA gene. The mutant genes were expressed in vivo using an inducible tac promoter. Mutant species of 23 S rRNA, containing deletions between positions 40 and 2773, were incorporated into stable ribonucleoprotein particles. In contrast, if one end of the 23 S rRNA was deleted, the mutant rRNA was unstable and did not form ribosomal particles. Protein composition of the mutant particles was specific; the presence of the primary rRNA-binding proteins corresponded to their known binding sites. Furthermore, several previously unknown ribosomal protein binding sites in 23 S rRNA were identified. Implications of the results on ribosome assembly are discussed.
authors.affiliation: Department of Molecular Biology, Tartu University, Estonia.
authors.forename: A
authors.initials: A
authors.lastname: Liiv
authors.forename: T
authors.initials: T
authors.lastname: Tenson
authors.forename: J
authors.initials: J
authors.lastname: Remme
mesh_terms: D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000096:biosynthesis / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000032:analysis / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry*; D017384:Sequence Deletion
chemical_list: D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; D012269:Ribosomal Proteins