title: The ribosome biogenesis factor yUtp23/hUTP23 coordinates key interactions in the yeast and human pre-40S particle and hUTP23 contains an essential PIN domain.
abstract: Two proteins with PIN endonuclease domains, yUtp24(Fcf1)/hUTP24 and yUtp23/hUTP23 are essential for early pre-ribosomal (r)RNA cleavages at sites A0, A1/1 and A2/2a in yeast and humans. The yUtp24/hUTP24 PIN endonuclease is proposed to cleave at sites A1/1 and A2/2a, but the enzyme cleaving at site A0 is not known. Yeast yUtp23 contains a degenerate, non-essential PIN domain and functions together with the snR30 snoRNA, while human hUTP23 is associated with U17, the human snR30 counterpart. Using in vivo RNA-protein crosslinking and gel shift experiments, we reveal that yUtp23/hUTP23 makes direct contacts with expansion sequence 6 (ES6) in the 18S rRNA sequence and that yUtp23 interacts with the 3΄ half of the snR30 snoRNA. Protein-protein interaction studies further demonstrated that yeast yUtp23 and human hUTP23 directly interact with the H/ACA snoRNP protein yNhp2/hNHP2, the RNA helicase yRok1/hROK1(DDX52), the ribosome biogenesis factor yRrp7/hRRP7 and yUtp24/hUTP24. yUtp23/hUTP23 could therefore be central to the coordinated integration and release of ES6 binding factors and likely plays a pivotal role in remodeling this pre-rRNA region in both yeast and humans. Finally, studies using RNAi-rescue systems in human cells revealed that intact PIN domain and Zinc finger motifs in human hUTP23 are essential for 18S rRNA maturation.
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
authors.forename: Graeme R
authors.initials: GR
authors.lastname: Wells
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
authors.forename: Franziska
authors.initials: F
authors.lastname: Weichmann
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
authors.forename: Katherine E
authors.initials: KE
authors.lastname: Sloan
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
authors.forename: David
authors.initials: D
authors.lastname: Colvin
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Watkins
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
authors.forename: Claudia
authors.initials: C
authors.lastname: Schneider
mesh_terms: D006801:Humans; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D000072417:Protein Domains / Q000235:genetics; D060066:Protein Interaction Maps / Q000235:genetics; D012322:RNA Precursors / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000737:chemistry / Q000235:genetics; D020537:RNA, Small Nucleolar / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D020718:Ribonucleoproteins, Small Nucleolar / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012270:Ribosomes / Q000737:chemistry / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics
chemical_list: D009687:Nuclear Proteins; D012322:RNA Precursors; D012337:RNA, Ribosomal, 18S; D020537:RNA, Small Nucleolar; D020718:Ribonucleoproteins, Small Nucleolar; D029701:Saccharomyces cerevisiae Proteins; C573244:Utp23 protein, S cerevisiae