title: Nucleolar proteins Bfr2 and Enp2 interact with DEAD-box RNA helicase Dbp4 in two different complexes.
abstract: Different pre-ribosomal complexes are formed during ribosome biogenesis, and the composition of these complexes is highly dynamic. Dbp4, a conserved DEAD-box RNA helicase implicated in ribosome biogenesis, interacts with nucleolar proteins Bfr2 and Enp2. We show that, like Dbp4, Bfr2 and Enp2 are required for the early processing steps leading to the production of 18S ribosomal RNA. We also found that Bfr2 and Enp2 associate with the U3 small nucleolar RNA (snoRNA), the U3-specific protein Mpp10 and various pre-18S ribosomal RNA species. Thus, we propose that Bfr2, Dbp4 and Enp2 are components of the small subunit (SSU) processome, a large complex of ∼80S. Sucrose gradient sedimentation analyses indicated that Dbp4, Bfr2 and Enp2 sediment in a peak of ∼50S and in a peak of ∼80S. Bfr2, Dbp4 and Enp2 associate together in the 50S complex, which does not include the U3 snoRNA; however, they associate with U3 snoRNA in the 80S complex (SSU processome). Immunoprecipitation experiments revealed that U14 snoRNA associates with Dbp4 in the 50S complex, but not with Bfr2 or Enp2. The assembly factor Tsr1 is not part of the '50S' complex, indicating this complex is not a pre-40S ribosome. A combination of experiments leads us to propose that Bfr2, Enp2 and Dbp4 are recruited at late steps during assembly of the SSU processome.
authors.affiliation: Département des sciences biologiques and Centre de recherche BioMed, Université du Québec à Montréal, Montréal, Québec, Canada.
authors.forename: Sahar
authors.initials: S
authors.lastname: Soltanieh
authors.forename: Martin
authors.initials: M
authors.lastname: Lapensée
authors.forename: François
authors.initials: F
authors.lastname: Dragon
mesh_terms: D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D012316:RNA Nucleotidyltransferases / Q000378:metabolism*; D012322:RNA Precursors / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012337:RNA, Ribosomal, 18S / Q000378:metabolism; D020537:RNA, Small Nucleolar / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: C069491:BFR2 protein, S cerevisiae; C111098:MPP10 protein, S cerevisiae; D009687:Nuclear Proteins; D010750:Phosphoproteins; D012322:RNA Precursors; D012337:RNA, Ribosomal, 18S; D020537:RNA, Small Nucleolar; C117378:RNA, U3 small nucleolar; D012261:Ribonucleoproteins; D029701:Saccharomyces cerevisiae Proteins; D012316:RNA Nucleotidyltransferases; C106767:HCA4 protein, S cerevisiae; D053487:DEAD-box RNA Helicases