title: Transcriptional repressor NIR functions in the ribosome RNA processing of both 40S and 60S subunits.
abstract: BACKGROUND
  NIR was identified as an inhibitor of histone acetyltransferase and it represses transcriptional activation of p53. NIR is predominantly localized in the nucleolus and known as Noc2p, which is involved in the maturation of the 60S ribosomal subunit. However, how NIR functions in the nucleolus remains undetermined. In the nucleolus, a 47S ribosomal RNA precursor (pre-rRNA) is transcribed and processed to produce 18S, 5.8S and 28S rRNAs. The 18S rRNA is incorporated into the 40S ribosomal subunit, whereas the 28S and 5.8S rRNAs are incorporated into the 60S subunit. U3 small nucleolar RNA (snoRNA) directs 18S rRNA processing and U8 snoRNA mediates processing of 28S and 5.8 S rRNAs. Functional disruption of nucleolus often causes p53 activation to inhibit cell proliferation.
  
  
  RESULTS
  Western blotting showed that NIR is ubiquitously expressed in different human cell lines. Knock-down of NIR by siRNA led to inhibition of the 18S, 28S and 5.8S rRNAs evaluated by pulse-chase experiment. Pre-rRNA particles (pre-rRNPs) were fractionated from the nucleus by sucrose gradient centrifugation and analysis of the pre-RNPs components showed that NIR existed in the pre-RNPs of both the 60S and 40S subunits and co-fractionated with 32S and 12S pre-rRNAs in the 60S pre-rRNP. Protein-RNA binding experiments demonstrated that NIR is associated with the 32S pre-rRNA and U8 snoRNA. In addition, NIR bound U3 snoRNA. It is a novel finding that depletion of NIR did not affect p53 protein level but de-repressed acetylation of p53 and activated p21.
  
  
  CONCLUSIONS
  We provide the first evidence for a transcriptional repressor to function in the rRNA biogenesis of both the 40S and 60S subunits. Our findings also suggested that a nucleolar protein may alternatively signal to p53 by affecting the p53 modification rather than affecting p53 protein level.
authors.affiliation: Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Peking University School of Oncology, Beijing Cancer Hospital & Institute, Beijing, China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Wu
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yingshuang
authors.initials: Y
authors.lastname: Wang
authors.forename: Ruirui
authors.initials: R
authors.lastname: Kong
authors.forename: Lelin
authors.initials: L
authors.lastname: Hu
authors.forename: Roland
authors.initials: R
authors.lastname: Schuele
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Du
authors.forename: Yang
authors.initials: Y
authors.lastname: Ke
mesh_terms: D000107:Acetylation; D002460:Cell Line; D002466:Cell Nucleolus / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D011485:Protein Binding; D021381:Protein Transport; D012322:RNA Precursors / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*; D012335:RNA, Ribosomal / Q000378:metabolism; D020537:RNA, Small Nucleolar / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism; D054683:Ribosome Subunits, Large, Eukaryotic / Q000378:metabolism*; D054682:Ribosome Subunits, Small, Eukaryotic / Q000378:metabolism*; D014158:Transcription, Genetic; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: C507220:NOC2L protein, human; D012322:RNA Precursors; D012335:RNA, Ribosomal; D020537:RNA, Small Nucleolar; C117378:RNA, U3 small nucleolar; D012097:Repressor Proteins; D012261:Ribonucleoproteins; D016159:Tumor Suppressor Protein p53