title: The RNA recognition motif of NIFK is required for rRNA maturation during cell cycle progression.
abstract: Ribosome biogenesis governs protein synthesis. NIFK is transactivated by c-Myc, the key regulator of ribosome biogenesis. The biological function of human NIFK is not well established, except that it has been shown to interact with Ki67 and NPM1. Here we report that NIFK is required for cell cycle progression and participates in the ribosome biogenesis via its RNA recognition motif (RRM). We show that silencing of NIFK inhibits cell proliferation through a reversible p53-dependent G1 arrest, possibly by induction of the RPL5/RPL11-mediated nucleolar stress. Mechanistically it is the consequence of impaired maturation of 28S and 5.8S rRNA resulting from inefficient cleavage of internal transcribed spacer (ITS) 1, a critical step in the separation of pre-ribosome to small and large subunits. Complementation of NIFK silencing by mutants shows that RNA-binding ability of RRM is essential for the pre-rRNA processing and G1 progression. More specifically, we validate that the RRM of NIFK preferentially binds to the 5'-region of ITS2 rRNA likely in both sequence specific and secondary structure dependent manners. Our results show how NIFK is involved in cell cycle progression through RRM-dependent pre-rRNA maturation, which could enhance our understanding of the function of NIFK in cell proliferation, and potentially also cancer and ribosomopathies.
authors.affiliation: a Institute of Biological Chemistry; Academia Sinica ; Taipei , Taiwan.
authors.forename: Wen-An
authors.initials: WA
authors.lastname: Pan
authors.forename: Hsin-Yue
authors.initials: HY
authors.lastname: Tsai
authors.forename: Shun-Chang
authors.initials: SC
authors.lastname: Wang
authors.forename: Michael
authors.initials: M
authors.lastname: Hsiao
authors.forename: Pei-Yu
authors.initials: PY
authors.lastname: Wu
authors.forename: Ming-Daw
authors.initials: MD
authors.lastname: Tsai
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D021903:DNA, Ribosomal Spacer / Q000235:genetics / Q000378:metabolism; D059585:G1 Phase Cell Cycle Checkpoints / Q000235:genetics; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D009690:Nucleic Acid Conformation; D000090243:Nucleophosmin; D059372:Nucleotide Motifs; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D011485:Protein Binding; D014176:Protein Biosynthesis; D012322:RNA Precursors / Q000235:genetics* / Q000378:metabolism; D012339:RNA, Ribosomal, 28S / Q000235:genetics* / Q000378:metabolism; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D015398:Signal Transduction; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D021903:DNA, Ribosomal Spacer; D047908:Intracellular Signaling Peptides and Proteins; C492406:NIFK protein, human; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D012322:RNA Precursors; D012339:RNA, Ribosomal, 28S; D012340:RNA, Ribosomal, 5.8S; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D000090243:Nucleophosmin
keywords: 5S RNP, 5S ribonucleoprotein particle; CDK1, cyclin dependent kinase 1; DFC, dense fibrillar component; ETS/ITS, external/internal transcribed spacers; GSK3, glycogen synthase kinase 3; Ki67; Ki67FHAID, Ki67-FHA interaction domain; LSU, large subunit; MDM2, murine double minute 2; NIFK, Nucleolar protein Interacting with the FHA domain of pKi-67; NPM1/B23, nucleophosmin; Noprecipitation; PAR-CLIP, Photo-Activatable-Ribonucleoside-Enhanced Crosslinking and Immu-pre-rRNAs, rRNA precursors; REMSA, RNA electrophoresis mobility shift assay; RNA recognition motif; RNP1 and 2, ribonucleoprotein motif 1 and 2; RPL5 and RPL11, large ribosomal protein 5 and 11; RRM, RNA recognition motif; cell cycle; nucleolar stress; rNIFK, recombinant NIFK; ribosome biogenesis; snoRNP, small nucleolar ribonucleoprotein
references.citation: EMBO J. 2003 Dec 15;22(24):6573-83
references.pmid: 14657029
references.citation: Mol Cell Biol. 1998 Mar;18(3):1181-9
references.pmid: 9488433
references.citation: Eur J Biochem. 1993 Feb 15;212(1):211-5
references.pmid: 8444156
references.citation: Cell. 1995 Aug 25;82(4):675-84
references.pmid: 7664346
references.citation: Cancer Res. 1995 Nov 15;55(22):5187-90
references.pmid: 7585571
references.citation: Stem Cells. 2010 Sep;28(9):1487-97
references.pmid: 20629179
references.citation: Nucleic Acids Res. 2011 Jan;39(1):280-91
references.pmid: 20805244
references.citation: Nucleic Acids Res. 2011 Mar;39(5):1811-22
references.pmid: 21036871
references.citation: Semin Hematol. 2011 Apr;48(2):97-105
references.pmid: 21435506
references.citation: EMBO J. 2011 Oct 5;30(19):4006-19
references.pmid: 21811236
references.citation: EMBO J. 2011 Oct 5;30(19):4020-32
references.pmid: 21926967
references.citation: Biochimie. 2012 Jul;94(7):1521-32
references.pmid: 22342225
references.citation: Cell. 2012 Aug 31;150(5):1068-81
references.pmid: 22939629
references.citation: J Cell Biol. 2013 Mar 4;200(5):577-88
references.pmid: 23439679
references.citation: Nucleic Acids Res. 2013 Jul;41(Web Server issue):W471-4
references.pmid: 23620284
references.citation: Cell Rep. 2013 Jul 11;4(1):87-98
references.pmid: 23831031
references.citation: Mol Cell. 2013 Aug 22;51(4):539-51
references.pmid: 23973377
references.citation: Cell Rep. 2013 Oct 17;5(1):237-47
references.pmid: 24120868
references.citation: Elife. 2014;3:e01641
references.pmid: 24867636
references.citation: J Cell Sci. 1996 Jun;109 ( Pt 6):1253-63
references.pmid: 8799815
references.citation: Mol Cell Biol. 2010 Sep;30(18):4404-14
references.pmid: 20647540
references.citation: Nucleic Acids Res. 1998 Oct 1;26(19):4508-15
references.pmid: 9742256
references.citation: Nucleic Acids Res. 1999 Jun 1;27(11):2271-82
references.pmid: 10325414
references.citation: FEBS J. 2005 May;272(9):2118-31
references.pmid: 15853797
references.citation: Nat Struct Mol Biol. 2005 Nov;12(11):987-93
references.pmid: 16244663
references.citation: Nucleic Acids Res. 2006;34(10):3030-43
references.pmid: 16738141
references.citation: Nat Rev Mol Cell Biol. 2007 Jul;8(7):574-85
references.pmid: 17519961
references.citation: Curr Protoc Mol Biol. 2004 Sep;Chapter 4:Unit 4.9
references.pmid: 18265351
references.citation: Curr Protoc Cell Biol. 2008 Jun;Chapter 22:Unit 22.11
references.pmid: 18551418
references.citation: Am J Pathol. 2008 Aug;173(2):301-10
references.pmid: 18583314
references.citation: Cell Mol Life Sci. 2008 Aug;65(15):2334-59
references.pmid: 18408888
references.citation: Nat Protoc. 2008;3(8):1312-7
references.pmid: 18714299
references.citation: PLoS One. 2008;3(8):e2987
references.pmid: 18714337
references.citation: Nature. 2008 Dec 18;456(7224):971-5
references.pmid: 19011615
references.citation: Nat Cell Biol. 2009 Apr;11(4):501-8
references.pmid: 19287375
references.citation: Proc Natl Acad Sci U S A. 2009 Jun 16;106(24):9613-8
references.pmid: 19482942
references.citation: Br J Haematol. 2009 Jun;145(6):806-15
references.pmid: 19438500
references.citation: Cancer Cell. 2009 Nov 6;16(5):369-77
references.pmid: 19878869
references.citation: Nat Rev Cancer. 2010 Apr;10(4):301-9
references.pmid: 20332779
references.citation: Nat Rev Cancer. 2003 Mar;3(3):179-92
references.pmid: 12612653
references.citation: Nucleic Acids Res. 2003 Nov 1;31(21):6148-56
references.pmid: 14576301
references.citation: EMBO J. 2003 Nov 17;22(22):6068-77
references.pmid: 14609953
references.citation: FEBS Lett. 2014 Aug 19;588(16):2571-9
references.pmid: 24747423
references.citation: J Biol Chem. 2014 Aug 15;289(33):22877-87
references.pmid: 25012651
references.citation: Nucleic Acids Res. 1999 Dec 1;27(23):4533-40
references.pmid: 10556307
references.citation: J Cell Physiol. 2000 Mar;182(3):311-22
references.pmid: 10653597
references.citation: Mol Cell Biol. 2000 Aug;20(15):5516-28
references.pmid: 10891491
references.citation: Mol Cell Biol. 2000 Aug;20(16):5930-8
references.pmid: 10913176
references.citation: Nat Struct Biol. 2001 Feb;8(2):141-5
references.pmid: 11175903
references.citation: Mol Cell Biol. 2001 Jul;21(13):4246-55
references.pmid: 11390653
references.citation: J Biol Chem. 2001 Jul 6;276(27):25386-91
references.pmid: 11342549
references.citation: Cell. 2010 Apr 2;141(1):129-41
references.pmid: 20371350
references.citation: J Biol Chem. 2010 Apr 16;285(16):12416-25
references.pmid: 20159984
references.citation: Cell Res. 2010 Jun;20(6):614-21
references.pmid: 20404858
references.citation: J Immunol. 1984 Oct;133(4):1710-5
references.pmid: 6206131