Reversible phosphorylation of Rpn1 regulates 26S proteasome assembly and function.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
Read More Add to Saved list
  • Additional Information
    • Source:
      Publisher: National Academy of Sciences Country of Publication: United States NLM ID: 7505876 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1091-6490 (Electronic) Linking ISSN: 00278424 NLM ISO Abbreviation: Proc Natl Acad Sci U S A Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, DC : National Academy of Sciences
    • Subject Terms:
      Membrane Proteins/*metabolism ; Nuclear Proteins/*metabolism ; Phosphoprotein Phosphatases/*metabolism ; Proteasome Endopeptidase Complex/*metabolism ; Protein Serine-Threonine Kinases/*metabolism ; Protein Subunits/*metabolism; Animals ; CRISPR-Cas Systems/genetics ; Cell Line ; Enzyme Assays ; Gene Knock-In Techniques ; Humans ; Membrane Proteins/genetics ; Mice ; Mice, Knockout ; Mice, Transgenic ; Mitochondria/metabolism ; Nuclear Proteins/genetics ; Oxidative Stress ; Phosphoprotein Phosphatases/genetics ; Phosphorylation/physiology ; Proteasome Endopeptidase Complex/genetics ; Protein Serine-Threonine Kinases/genetics ; Protein Subunits/genetics ; RNA, Small Interfering/metabolism ; Serine/metabolism ; Trans-Activators/genetics ; Trans-Activators/metabolism
    • Abstract:
      The fundamental importance of the 26S proteasome in health and disease suggests that its function must be finely controlled, and yet our knowledge about proteasome regulation remains limited. Posttranslational modifications, especially phosphorylation, of proteasome subunits have been shown to impact proteasome function through different mechanisms, although the vast majority of proteasome phosphorylation events have not been studied. Here, we have characterized 1 of the most frequently detected proteasome phosphosites, namely Ser361 of Rpn1, a base subunit of the 19S regulatory particle. Using a variety of approaches including CRISPR/Cas9-mediated gene editing and quantitative mass spectrometry, we found that loss of Rpn1-S361 phosphorylation reduces proteasome activity, impairs cell proliferation, and causes oxidative stress as well as mitochondrial dysfunction. A screen of the human kinome identified several kinases including PIM1/2/3 that catalyze S361 phosphorylation, while its level is reversibly controlled by the proteasome-resident phosphatase, UBLCP1. Mechanistically, Rpn1-S361 phosphorylation is required for proper assembly of the 26S proteasome, and we have utilized a genetic code expansion system to directly demonstrate that S361-phosphorylated Rpn1 more readily forms a precursor complex with Rpt2, 1 of the first steps of 19S base assembly. These findings have revealed a prevalent and biologically important mechanism governing proteasome formation and function.
      Competing Interests: The authors declare no competing interest.
    • References:
      Science. 2016 Feb 19;351(6275):null. (PMID: 26912900)
      J Mol Biol. 2017 Nov 10;429(22):3500-3524. (PMID: 28583440)
      Protein Cell. 2017 Apr;8(4):255-272. (PMID: 28258412)
      Redox Biol. 2017 Oct;13:550-567. (PMID: 28763764)
      Science. 2001 May 25;292(5521):1552-5. (PMID: 11375494)
      Nat Rev Cancer. 2011 Jan;11(1):23-34. (PMID: 21150935)
      Cell Rep. 2014 Jun 12;7(5):1371-1380. (PMID: 24857655)
      Nat Rev Drug Discov. 2018 Sep;17(9):660-688. (PMID: 30116051)
      J Biol Chem. 2017 Sep 29;292(39):16310-16320. (PMID: 28821611)
      J Biol Chem. 2009 Sep 25;284(39):26655-65. (PMID: 19638347)
      Nat Rev Mol Cell Biol. 2018 Nov;19(11):697-712. (PMID: 30065390)
      Bioorg Med Chem. 2015 Jun 15;23(12):2798-809. (PMID: 25907364)
      Cell. 2009 May 29;137(5):914-25. (PMID: 19490896)
      J Biol Chem. 1998 Dec 25;273(52):34970-5. (PMID: 9857028)
      Biochim Biophys Acta. 2011 Feb;1809(2):80-7. (PMID: 20674813)
      Biochem Biophys Res Commun. 1995 Aug 4;213(1):7-14. (PMID: 7639764)
      J Biol Chem. 2000 Jan 14;275(2):875-82. (PMID: 10625621)
      Biochem J. 2017 Sep 24;474(19):3355-3371. (PMID: 28947610)
      Annu Rev Biochem. 2018 Jun 20;87:697-724. (PMID: 29652515)
      Proc Natl Acad Sci U S A. 2011 Nov 15;108(46):18649-54. (PMID: 21949367)
      Nature. 2019 Jan;565(7737):49-55. (PMID: 30479383)
      Cell. 2009 May 29;137(5):887-99. (PMID: 19446322)
      Trends Biochem Sci. 2017 Sep;42(9):712-725. (PMID: 28579074)
      Nat Cell Biol. 2016 Feb;18(2):202-12. (PMID: 26655835)
      Nat Struct Mol Biol. 2016 Sep;23(9):778-85. (PMID: 27428775)
      Proc Natl Acad Sci U S A. 2018 Aug 7;115(32):8155-8160. (PMID: 29987021)
      Nature. 2009 Jun 11;459(7248):861-5. (PMID: 19412159)
      Nat Rev Mol Cell Biol. 2009 Feb;10(2):104-15. (PMID: 19165213)
      J Clin Invest. 2005 Oct;115(10):2618-24. (PMID: 16200194)
      Nature. 2012 Sep 13;489(7415):263-8. (PMID: 22922647)
      Nat Commun. 2015 Sep 09;6:8130. (PMID: 26350500)
      Cell Res. 2011 Jun;21(6):867-83. (PMID: 21537343)
      Biochim Biophys Acta. 2014 Jan;1843(1):13-25. (PMID: 23994620)
      Nature. 2009 Mar 26;458(7237):438-44. (PMID: 19325623)
      BMC Biol. 2014 Nov 11;12:94. (PMID: 25385277)
      Science. 2011 Aug 26;333(6046):1151-4. (PMID: 21868676)
      Cell. 2010 Dec 23;143(7):1174-89. (PMID: 21183079)
      Cell Stem Cell. 2012 Dec 7;11(6):783-98. (PMID: 23103054)
      Nature. 2013 Apr 18;496(7445):372-6. (PMID: 23503661)
      Science. 2018 Nov 30;362(6418):. (PMID: 30309908)
      Mol Cell Proteomics. 2012 Dec;11(12):1640-51. (PMID: 22942356)
      Nature. 2012 Sep 13;489(7415):304-8. (PMID: 22972301)
      Mol Cell Proteomics. 2017 May;16(5):840-854. (PMID: 28292943)
      Biochemistry. 2001 Jan 16;40(2):314-9. (PMID: 11148024)
      Sci Signal. 2017 Apr 11;10(474):null. (PMID: 28400531)
      Nature. 2009 Jun 11;459(7248):866-70. (PMID: 19412160)
      Curr Biol. 2018 Feb 19;28(4):R170-R185. (PMID: 29462587)
      Redox Biol. 2017 Oct;13:452-458. (PMID: 28715730)
      Biochem Biophys Res Commun. 2016 Apr 22;473(1):80-86. (PMID: 26996128)
      Cell Metab. 2014 Apr 1;19(4):642-52. (PMID: 24703696)
      Mol Cell. 2009 Oct 23;36(2):326-39. (PMID: 19854140)
      Autophagy. 2018;14(8):1293-1309. (PMID: 29909722)
      Proc Natl Acad Sci U S A. 2015 Dec 29;112(52):E7176-85. (PMID: 26669444)
      Aging Cell. 2018 Oct;17(5):e12801. (PMID: 29992725)
      Open Biol. 2017 May;7(5):. (PMID: 28539385)
      Cell. 2012 Aug 31;150(5):987-1001. (PMID: 22939624)
      Nat Med. 2016 Jan;22(1):46-53. (PMID: 26692334)
      Annu Rev Biochem. 2009;78:477-513. (PMID: 19489727)
      Annu Rev Biochem. 1998;67:425-79. (PMID: 9759494)
      Cell. 2010 Feb 19;140(4):567-78. (PMID: 20178748)
      Cell. 2017 May 18;169(5):792-806. (PMID: 28525752)
      Mol Cell. 2017 Jul 20;67(2):322-333.e6. (PMID: 28689658)
    • Grant Information:
      R01 DK018024 United States DK NIDDK NIH HHS; R01 GM074830 United States GM NIGMS NIH HHS
    • Contributed Indexing:
      Keywords: PIM; UBLCP1; genetic code expansion; phosphorylation; proteasome
    • Accession Number:
      0 (Membrane Proteins)
      0 (Nuclear Proteins)
      0 (PSMD14 protein, mouse)
      0 (Protein Subunits)
      0 (RNA, Small Interfering)
      0 (RPN1 protein, human)
      0 (Trans-Activators)
      0 (ribophorin)
      452VLY9402 (Serine)
      EC 2.7.11.1 (Protein Serine-Threonine Kinases)
      EC 3.1.3.16 (Phosphoprotein Phosphatases)
      EC 3.1.3.16 (UBLCP1 protein, human)
      EC 3.4.25.1 (Proteasome Endopeptidase Complex)
      EC 3.4.99.- (ATP dependent 26S protease)
    • Publication Date:
      Date Created: 20191218 Date Completed: 20200511 Latest Revision: 20211204
    • Publication Date:
      20231215
    • Accession Number:
      PMC6955308
    • Accession Number:
      10.1073/pnas.1912531117
    • Accession Number:
      31843888