Home LiteratureArticle Details
PMID: 18716056 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Lysosomal localization of ubiquitinated Jun requires multiple determinants in a lysine-27-linked polyubiquitin conjugate.

Molecular biology of the cell ·Vol. 19 ·No. 11 ·2008-11-00 ·Pages 4588-601

Ikeda H, Kerppola TK

Abstract

Ubiquitination regulates many cellular functions, including protein localization and degradation. Each function is specified by unique determinants in the conjugate. Ubiquitinated Jun is localized to lysosomes for degradation. Here, we characterized determinants of Jun ubiquitination and lysosomal localization by using ubiquitin-mediated fluorescence complementation (UbFC) in living cells and analysis of the stoichiometry of ubiquitin linked to Jun extracted from cells. The delta region of Jun and isoleucine-44 in ubiquitin were required for lysosomal localization of the conjugate. Ubiquitin containing only lysine-27, but no other single-lysine ubiquitin, mediated Jun ubiquitination, albeit at lower stoichiometry than wild-type ubiquitin. These conjugates were predominantly nuclear, but coexpression of lysine-27 and lysine-less ubiquitins enhanced the mean stoichiometry of Jun ubiquitination and lysosomal localization of the conjugate. Hepatocyte growth factor-regulated tyrosine kinase substrate (HRS) and tumor susceptibility gene 101 (TSG101) colocalized with ubiquitinated Jun. Knockdown of HRS or TSG101 inhibited lysosomal localization of ubiquitinated Jun and reduced Jun turnover. Ubiquitination of other Fos and Jun family proteins had distinct effects on their localization. Our results indicate that Jun is polyubiquitinated by E3 ligases that produce lysine-27-linked chains. Lysosomal localization of the conjugate requires determinants in Jun and in ubiquitin that are recognized in part by TSG101 and HRS, facilitating selective translocation and degradation of ubiquitinated Jun.

MeSH Terms
Animals COS Cells Chlorocebus aethiops DNA-Binding Proteins/metabolism Endocytosis Endosomal Sorting Complexes Required for Transport ErbB Receptors/metabolism Fluorescence Humans Lysine/metabolism Lysosomes/metabolism Mutant Proteins/metabolism Phosphoproteins/metabolism Polyubiquitin/metabolism Protein Processing, Post-Translational Protein Transport Proto-Oncogene Proteins c-fos/metabolism Proto-Oncogene Proteins c-jun/genetics,metabolism Transcription Factors/metabolism Transcriptional Activation Ubiquitination
Chemicals
DNA-Binding Proteins Endosomal Sorting Complexes Required for Transport Mutant Proteins Phosphoproteins Proto-Oncogene Proteins c-fos Proto-Oncogene Proteins c-jun Transcription Factors Tsg101 protein hepatocyte growth factor-regulated tyrosine kinase substrate Polyubiquitin ErbB Receptors Lysine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ikeda Hiromi
Howard Hughes Medical Institute and Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109-0650, USA.
Kerppola Tom K
References (34)
34 references, click to expand
  1. Protein regulation by monoubiquitin.
    Nat Rev Mol Cell Biol. 2001 Mar;2(3):195-201 PMID: 11265249
  2. Multiple monoubiquitination of RTKs is sufficient for their endocytosis and degradation.
    Nat Cell Biol. 2003 May;5(5):461-6 PMID: 12717448
  3. Ubiquitin-dependent sorting into the multivesicular body pathway requires the function of a conserved endosomal protein sorting complex, ESCRT-I.
    Cell. 2001 Jul 27;106(2):145-55 PMID: 11511343
  4. Polyubiquitin chains: polymeric protein signals.
    Curr Opin Chem Biol. 2004 Dec;8(6):610-6 PMID: 15556404
  5. The tyrosine kinase c-Abl protects c-Jun from ubiquitination-mediated degradation in T cells.
    J Biol Chem. 2006 Oct 6;281(40):29711-8 PMID: 16901904
  6. Autophagy as a regulated pathway of cellular degradation.
    Science. 2000 Dec 1;290(5497):1717-21 PMID: 11099404
  7. Surface hydrophobic residues of multiubiquitin chains essential for proteolytic targeting.
    Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):861-6 PMID: 8570649
  8. A proteomics approach to understanding protein ubiquitination.
    Nat Biotechnol. 2003 Aug;21(8):921-6 PMID: 12872131
  9. Ubiquitin-dependent c-Jun degradation in vivo is mediated by the delta domain.
    Cell. 1994 Sep 9;78(5):787-98 PMID: 8087846
  10. Polyubiquitination of p53 by a ubiquitin ligase activity of p300.
    Science. 2003 Apr 11;300(5617):342-4 PMID: 12690203
  11. Jab1 antagonizes TGF-beta signaling by inducing Smad4 degradation.
    EMBO Rep. 2002 Feb;3(2):171-6 PMID: 11818334
  12. A new group of conserved coactivators that increase the specificity of AP-1 transcription factors.
    Nature. 1996 Oct 3;383(6599):453-7 PMID: 8837781
  13. Degradation of the cyclin-dependent-kinase inhibitor p27Kip1 is instigated by Jab1.
    Nature. 1999 Mar 11;398(6723):160-5 PMID: 10086358
  14. Human De-etiolated-1 regulates c-Jun by assembling a CUL4A ubiquitin ligase.
    Science. 2004 Feb 27;303(5662):1371-4 PMID: 14739464
  15. Ubiquitin-mediated fluorescence complementation reveals that Jun ubiquitinated by Itch/AIP4 is localized to lysosomes.
    Proc Natl Acad Sci U S A. 2004 Oct 12;101(41):14782-7 PMID: 15469925
  16. The ubiquitin conjugation system is required for ligand-induced endocytosis and degradation of the growth hormone receptor.
    EMBO J. 1996 Aug 1;15(15):3806-12 PMID: 8670885
  17. The ubiquitin ligase SCFFbw7 antagonizes apoptotic JNK signaling.
    Science. 2004 Feb 27;303(5662):1374-8 PMID: 14739463
  18. Differential regulation of EGF receptor internalization and degradation by multiubiquitination within the kinase domain.
    Mol Cell. 2006 Mar 17;21(6):737-48 PMID: 16543144
  19. Jun turnover is controlled through JNK-dependent phosphorylation of the E3 ligase Itch.
    Science. 2004 Oct 8;306(5694):271-5 PMID: 15358865
  20. Mono- versus polyubiquitination: differential control of p53 fate by Mdm2.
    Science. 2003 Dec 12;302(5652):1972-5 PMID: 14671306
  21. Ubiquitin-binding domains.
    Nat Rev Mol Cell Biol. 2005 Aug;6(8):610-21 PMID: 16064137
  22. Jab1/CSN5, a component of the COP9 signalosome, regulates transforming growth factor beta signaling by binding to Smad7 and promoting its degradation.
    Mol Cell Biol. 2004 Mar;24(6):2251-62 PMID: 14993265
  23. Itch/AIP4 mediates Deltex degradation through the formation of K29-linked polyubiquitin chains.
    EMBO Rep. 2006 Nov;7(11):1147-53 PMID: 17028573
  24. TSG101 interaction with HRS mediates endosomal trafficking and receptor down-regulation.
    Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7626-31 PMID: 12802020
  25. FOXO4 transcriptional activity is regulated by monoubiquitination and USP7/HAUSP.
    Nat Cell Biol. 2006 Oct;8(10):1064-73 PMID: 16964248
  26. Age-dependent accumulation of ubiquitinated 2',3'-cyclic nucleotide 3'-phosphodiesterase in myelin lipid rafts.
    Glia. 2008 Jan 1;56(1):118-33 PMID: 17963267
  27. The COP9 signalosome: at the interface between signal transduction and ubiquitin-dependent proteolysis.
    J Cell Sci. 2002 Feb 1;115(Pt 3):467-73 PMID: 11861754
  28. The ESCRT complexes: structure and mechanism of a membrane-trafficking network.
    Annu Rev Biophys Biomol Struct. 2006;35:277-98 PMID: 16689637
  29. The v-Jun point mutation allows c-Jun to escape GSK3-dependent recognition and destruction by the Fbw7 ubiquitin ligase.
    Cancer Cell. 2005 Jul;8(1):25-33 PMID: 16023596
  30. Mammalian class E vps proteins recognize ubiquitin and act in the removal of endosomal protein-ubiquitin conjugates.
    J Cell Biol. 2002 Apr 1;157(1):91-101 PMID: 11916981
  31. Endocytosis of receptor tyrosine kinases is driven by monoubiquitylation, not polyubiquitylation.
    J Biol Chem. 2003 Jun 13;278(24):21323-6 PMID: 12719435
  32. Ligand-dependent degradation of Smad3 by a ubiquitin ligase complex of ROC1 and associated proteins.
    Mol Biol Cell. 2001 May;12(5):1431-43 PMID: 11359933
  33. The ubiquitin system.
    Annu Rev Biochem. 1998;67:425-79 PMID: 9759494
  34. Growth factor-induced tyrosine phosphorylation of Hrs, a novel 115-kilodalton protein with a structurally conserved putative zinc finger domain.
    Mol Cell Biol. 1995 Nov;15(11):6213-21 PMID: 7565774
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1939-4586
DOI
10.1091/mbc.e08-05-0496
Published
2008-11-00
Epub
2008-00-20
Pages
4588-601
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC2575148
Subset
IM
Grants
NIGMS NIH HHS · R01 GM086213 · United States
Howard Hughes Medical Institute · United States
External Links
PubMed source DOI: 10.1091/mbc.e08-05-0496 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2