title: The central core region of yeast ribosomal protein L11 is important for subunit joining and translational fidelity.
abstract: Yeast ribosomal protein L11 is positioned at the intersubunit cleft of the large subunit central protuberance, forming an intersubunit bridge with the small subunit protein S18. Mutants were engineered in the central core region of L11 which interacts with Helix 84 of the 25S rRNA. Numerous mutants in this region conferred 60S subunit biogenesis defects. Specifically, many mutations of F96 and the A66D mutant promoted formation of halfmers as assayed by sucrose density ultracentrifugation. Halfmer formation was not due to deficiency in 60S subunit production, suggesting that the mutants affected subunit-joining. Chemical modification analyses indicated that the A66D mutant, but not the F96 mutants, promoted changes in 25S rRNA structure, suggesting at least two modalities for subunit joining defects. 25S rRNA structural changes were located both adjacent to A66D (in H84), and more distant (in H96-7). While none of the mutants significantly affected ribosome/tRNA binding constants, they did have strong effects on cellular growth at both high and low temperatures, in the presence of translational inhibitors, and promoted changes in translational fidelity. Two distinct mechanisms are proposed by which L11 mutants may affect subunit joining, and identification of the amino acids associated with each of these processes are presented. These findings may have implications for our understanding of multifaceted diseases such as Diamond--Blackfan anemia which have been linked in part with mutations in L11.
authors.affiliation: Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA.
authors.forename: Michael H J
authors.initials: MH
authors.lastname: Rhodin
authors.forename: Rasa
authors.initials: R
authors.lastname: Rakauskaitė
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Dinman
mesh_terms: D001665:Binding Sites / Q000235:genetics; D009154:Mutation; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D015202:Protein Engineering / Q000379:methods; D012335:RNA, Ribosomal / Q000737:chemistry / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; C525863:Rpl12 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C027624:ribosomal protein L11; C065039:RNA, ribosomal, 25S