title: Roles of Dim2 in ribosome assembly.
abstract: In eukaryotes, ribosome assembly requires hundreds of conserved essential proteins not present in the mature particle. Despite their importance, the function of most factors remains unknown. This is because protein deletion often affects the composition of the entire particle. Additionally, many proteins are present in assembling ribosomes for extended times, which makes it difficult to pinpoint their role to a particular step. Here we have combined classical yeast biochemistry with experiments using recombinant proteins and RNA to study the role of Dim2 and its interaction with Nob1, the nuclease that generates the 3'-end of 18 S rRNA. Analysis of Dim2 mutants in which the interaction with Nob1 is disrupted demonstrates that this interaction between Dim2 and Nob1 is essential for optimal growth, and RNA binding experiments show that Dim2 increases Nob1 RNA affinity. Furthermore, our data indicate that Dim2 helps regulate Nob1 cleavage activity at the 3'-end of 18 S rRNA, as point mutants where this interaction is abolished in vitro accumulate pre-ribosomes containing Nob1 and 20 S rRNA in vivo. Interestingly, the site of interaction with Nob1 is mapped to the canonical RNA binding surface of a KH-like domain in Dim2, providing another example where an RNA-binding domain can be repurposed for protein interactions.
authors.affiliation: Program in Chemical Biology, University of Michigan, Ann Arbor, Michigan 48109-1055, USA.
authors.forename: Heather A
authors.initials: HA
authors.lastname: Woolls
authors.forename: Allison C
authors.initials: AC
authors.lastname: Lamanna
authors.forename: Katrin
authors.initials: K
authors.lastname: Karbstein
mesh_terms: D001665:Binding Sites; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D017354:Point Mutation; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism*; D012337:RNA, Ribosomal, 18S / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C484364:DIM2 protein, S cerevisiae; C405962:NOB1 protein, S cerevisiae; D009687:Nuclear Proteins; D012331:RNA, Fungal; D012337:RNA, Ribosomal, 18S; D029701:Saccharomyces cerevisiae Proteins