Home LiteratureArticle Details
PMID: 21172657 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Ubiquitylation of an ERAD substrate occurs on multiple types of amino acids.

Molecular cell ·Vol. 40 ·No. 6 ·2010-12-22 ·Pages 917-26

Shimizu Y, Okuda-Shimizu Y, Hendershot LM

Abstract

Any protein synthesized in the secretory pathway has the potential to misfold and would need to be recognized and ubiquitylated for degradation. This is astounding, since only a few ERAD-specific E3 ligases have been identified. To begin to understand substrate recognition, we wished to map the ubiquitylation sites on the NS-1 nonsecreted immunoglobulin light chain, which is an ERAD substrate. Ubiquitin is usually attached to lysine residues and less frequently to the N terminus of proteins. In addition, several viral E3s have been identified that attach ubiquitin to cysteine or serine/threonine residues. Mutation of lysines, serines, and threonines in the NS-1 variable region was necessary to significantly reduce ubiquitylation and stabilize the protein. The Hrd1 E3 ligase was required to modify all three amino acids. Our studies argue that ubiquitylation of ER proteins relies on very different mechanisms of recognition and modification than those used to regulate biological processes.

MeSH Terms
Animals COS Cells Cell Line Chlorocebus aethiops Cysteine/metabolism Endoplasmic Reticulum/metabolism Humans Hydrogen-Ion Concentration Immunoglobulin kappa-Chains/metabolism Lysine/genetics,metabolism Mice NIH 3T3 Cells Protein Structure, Tertiary Protein Unfolding Saccharomyces cerevisiae Proteins Serine/genetics,metabolism Sodium Hydroxide/metabolism Substrate Specificity Threonine/genetics,metabolism Ubiquitin/metabolism Ubiquitin-Protein Ligases Ubiquitination
Chemicals
Immunoglobulin kappa-Chains Saccharomyces cerevisiae Proteins Ubiquitin Threonine Serine Sodium Hydroxide HRD1 protein, S cerevisiae Ubiquitin-Protein Ligases Lysine Cysteine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Shimizu Yuichiro
Department of Genetics and Tumor Cell Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Okuda-Shimizu Yuki
Hendershot Linda M
References (49)
49 references, click to expand
  1. Distinct ubiquitin-ligase complexes define convergent pathways for the degradation of ER proteins.
    Cell. 2006 Jul 28;126(2):361-73 PMID: 16873066
  2. The ubiquitin-domain protein HERP forms a complex with components of the endoplasmic reticulum associated degradation pathway.
    J Mol Biol. 2005 Dec 16;354(5):1021-7 PMID: 16289116
  3. Derlin-2 and Derlin-3 are regulated by the mammalian unfolded protein response and are required for ER-associated degradation.
    J Cell Biol. 2006 Jan 30;172(3):383-93 PMID: 16449189
  4. The variable domain of nonassembled Ig light chains determines both their half-life and binding to the chaperone BiP.
    Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1574-8 PMID: 9465057
  5. AAA-ATPase p97/Cdc48p, a cytosolic chaperone required for endoplasmic reticulum-associated protein degradation.
    Mol Cell Biol. 2002 Jan;22(2):626-34 PMID: 11756557
  6. N-terminal acetyltransferases and sequence requirements for N-terminal acetylation of eukaryotic proteins.
    J Mol Biol. 2003 Jan 24;325(4):595-622 PMID: 12507466
  7. Function of the p97-Ufd1-Npl4 complex in retrotranslocation from the ER to the cytosol: dual recognition of nonubiquitinated polypeptide segments and polyubiquitin chains.
    J Cell Biol. 2003 Jul 7;162(1):71-84 PMID: 12847084
  8. Ubiquitin: structures, functions, mechanisms.
    Biochim Biophys Acta. 2004 Nov 29;1695(1-3):55-72 PMID: 15571809
  9. Degradation of the bile salt export pump at endoplasmic reticulum in progressive familial intrahepatic cholestasis type II.
    Hepatology. 2008 Nov;48(5):1558-69 PMID: 18798335
  10. Intracellular degradation of the transport-impaired human PiZ alpha 1-antitrypsin variant. Biochemical mapping of the degradative event among compartments of the secretory pathway.
    J Biol Chem. 1990 Aug 15;265(23):14001-7 PMID: 2380201
  11. Quality control in the endoplasmic reticulum.
    Nat Rev Mol Cell Biol. 2003 Mar;4(3):181-91 PMID: 12612637
  12. ERAD: the long road to destruction.
    Nat Cell Biol. 2005 Aug;7(8):766-72 PMID: 16056268
  13. Use of modular substrates demonstrates mechanistic diversity and reveals differences in chaperone requirement of ERAD.
    J Biol Chem. 2003 Sep 19;278(38):35903-13 PMID: 12847107
  14. N-terminal ubiquitination: more protein substrates join in.
    Trends Cell Biol. 2004 Mar;14(3):103-6 PMID: 15055197
  15. Synthesis and sequence-specific proteolysis of hybrid proteins produced in Escherichia coli.
    Methods Enzymol. 1987;153:461-81 PMID: 3323806
  16. Mutant analysis links the translocon and BiP to retrograde protein transport for ER degradation.
    Nature. 1997 Aug 28;388(6645):891-5 PMID: 9278052
  17. SEL1L, the homologue of yeast Hrd3p, is involved in protein dislocation from the mammalian ER.
    J Cell Biol. 2006 Oct 23;175(2):261-70 PMID: 17043138
  18. Serine residues in the cytosolic tail of the T-cell antigen receptor alpha-chain mediate ubiquitination and endoplasmic reticulum-associated degradation of the unassembled protein.
    J Biol Chem. 2010 Jul 30;285(31):23916-24 PMID: 20519503
  19. Ubiquitination on nonlysine residues by a viral E3 ubiquitin ligase.
    Science. 2005 Jul 1;309(5731):127-30 PMID: 15994556
  20. HRD gene dependence of endoplasmic reticulum-associated degradation.
    Mol Biol Cell. 2000 May;11(5):1697-708 PMID: 10793145
  21. Cytosolic degradation of T-cell receptor alpha chains by the proteasome.
    J Biol Chem. 1997 Aug 15;272(33):20800-4 PMID: 9252404
  22. A conserved ubiquitin ligase of the nuclear envelope/endoplasmic reticulum that functions in both ER-associated and Matalpha2 repressor degradation.
    Genes Dev. 2001 Oct 15;15(20):2660-74 PMID: 11641273
  23. The activity of a human endoplasmic reticulum-associated degradation E3, gp78, requires its Cue domain, RING finger, and an E2-binding site.
    Proc Natl Acad Sci U S A. 2006 Jan 10;103(2):341-6 PMID: 16407162
  24. TEB4 is a C4HC3 RING finger-containing ubiquitin ligase of the endoplasmic reticulum.
    Biochem J. 2005 Jun 1;388(Pt 2):647-55 PMID: 15673284
  25. Proteasome-dependent endoplasmic reticulum-associated protein degradation: an unconventional route to a familiar fate.
    Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13797-801 PMID: 8943015
  26. Mechanisms underlying ubiquitination.
    Annu Rev Biochem. 2001;70:503-33 PMID: 11395416
  27. Ubiquitination of serine, threonine, or lysine residues on the cytoplasmic tail can induce ERAD of MHC-I by viral E3 ligase mK3.
    J Cell Biol. 2007 May 21;177(4):613-24 PMID: 17502423
  28. One step at a time: endoplasmic reticulum-associated degradation.
    Nat Rev Mol Cell Biol. 2008 Dec;9(12):944-57 PMID: 19002207
  29. The ER-bound RING finger protein 5 (RNF5/RMA1) causes degenerative myopathy in transgenic mice and is deregulated in inclusion body myositis.
    PLoS One. 2008 Feb 13;3(2):e1609 PMID: 18270596
  30. A membrane protein complex mediates retro-translocation from the ER lumen into the cytosol.
    Nature. 2004 Jun 24;429(6994):841-7 PMID: 15215856
  31. Mechanism and function of deubiquitinating enzymes.
    Biochim Biophys Acta. 2004 Nov 29;1695(1-3):189-207 PMID: 15571815
  32. Characterization of an ERAD pathway for nonglycosylated BiP substrates, which require Herp.
    Mol Cell. 2007 Nov 30;28(4):544-54 PMID: 18042451
  33. BiP and immunoglobulin light chain cooperate to control the folding of heavy chain and ensure the fidelity of immunoglobulin assembly.
    Mol Biol Cell. 1999 Jul;10(7):2209-19 PMID: 10397760
  34. The requirement for molecular chaperones during endoplasmic reticulum-associated protein degradation demonstrates that protein export and import are mechanistically distinct.
    J Biol Chem. 1999 Feb 5;274(6):3453-60 PMID: 9920890
  35. A membrane protein required for dislocation of misfolded proteins from the ER.
    Nature. 2004 Jun 24;429(6994):834-40 PMID: 15215855
  36. Protein ubiquitination involving an E1-E2-E3 enzyme ubiquitin thioester cascade.
    Nature. 1995 Jan 5;373(6509):81-3 PMID: 7800044
  37. Apoptosis induction by Bid requires unconventional ubiquitination and degradation of its N-terminal fragment.
    J Cell Biol. 2007 Dec 31;179(7):1453-66 PMID: 18166654
  38. Interaction of BiP with newly synthesized immunoglobulin light chain molecules: cycles of sequential binding and release.
    EMBO J. 1992 Apr;11(4):1573-81 PMID: 1563355
  39. Human HRD1 is an E3 ubiquitin ligase involved in degradation of proteins from the endoplasmic reticulum.
    J Biol Chem. 2004 Jan 30;279(5):3525-34 PMID: 14593114
  40. A ubiquitin ligase transfers preformed polyubiquitin chains from a conjugating enzyme to a substrate.
    Nature. 2007 Mar 15;446(7133):333-7 PMID: 17310145
  41. Der1, a novel protein specifically required for endoplasmic reticulum degradation in yeast.
    EMBO J. 1996 Feb 15;15(4):753-63 PMID: 8631297
  42. The tumor autocrine motility factor receptor, gp78, is a ubiquitin protein ligase implicated in degradation from the endoplasmic reticulum.
    Proc Natl Acad Sci U S A. 2001 Dec 4;98(25):14422-7 PMID: 11724934
  43. Multiple lysine mutations in the C-terminal domain of p53 interfere with MDM2-dependent protein degradation and ubiquitination.
    Mol Cell Biol. 2000 Dec;20(24):9391-8 PMID: 11094089
  44. ER degradation of a misfolded luminal protein by the cytosolic ubiquitin-proteasome pathway.
    Science. 1996 Sep 20;273(5282):1725-8 PMID: 8781238
  45. Herp, a new ubiquitin-like membrane protein induced by endoplasmic reticulum stress.
    J Biol Chem. 2000 Oct 20;275(42):32846-53 PMID: 10922362
  46. EDEM1 recognition and delivery of misfolded proteins to the SEL1L-containing ERAD complex.
    Mol Cell. 2009 Jun 12;34(5):627-33 PMID: 19524542
  47. Ube2j2 ubiquitinates hydroxylated amino acids on ER-associated degradation substrates.
    J Cell Biol. 2009 Nov 30;187(5):655-68 PMID: 19951915
  48. The E3 ubiquitin ligase TEB4 mediates degradation of type 2 iodothyronine deiodinase.
    Mol Cell Biol. 2009 Oct;29(19):5339-47 PMID: 19651899
  49. The human cytomegalovirus US11 gene product dislocates MHC class I heavy chains from the endoplasmic reticulum to the cytosol.
    Cell. 1996 Mar 8;84(5):769-79 PMID: 8625414
Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-4164
DOI
10.1016/j.molcel.2010.11.033
Published
2010-12-22
Pages
917-26
Language
English
Region
United States
NLM ID
9802571
PMCID
PMC3031134
Subset
IM
Grants
NIGMS NIH HHS · GM54068 · United States
NCI NIH HHS · CA21765 · United States
NCI NIH HHS · P30 CA021765 · United States
NIGMS NIH HHS · R01 GM054068-15 · United States
NIGMS NIH HHS · R01 GM054068 · United States
External Links
PubMed source DOI: 10.1016/j.molcel.2010.11.033 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: [email protected]

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