Home LiteratureArticle Details
PMID: 22212814 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Sna3 is an Rsp5 adaptor protein that relies on ubiquitination for its MVB sorting.

Traffic (Copenhagen, Denmark) ·Vol. 13 ·No. 4 ·2012-04-00 ·Pages 586-98

MacDonald C, Stringer DK, Piper RC

Abstract

The process in which ubiquitin (Ub) conjugation is required for trafficking of integral membrane proteins into multivesicular bodies (MVBs) and eventual degradation in the lumen of lysosomes/vacuoles is well defined. However, Ub-independent pathways into MVBs are less understood. To better understand this process, we have further characterized the membrane protein Sna3, the prototypical Ub-independent cargo protein sorted through the MVB pathway in yeast. We show that Sna3 trafficking to the vacuole is critically dependent on Rsp5 ligase activity and ubiquitination. We find Sna3 undergoes Ub-dependent MVB sorting by either becoming ubiquitinated itself or associating with other ubiquitinated membrane protein substrates. In addition, our functional studies support a role for Sna3 as an adaptor protein that recruits Rsp5 to cargo such as the methionine transporter Mup1, resulting in efficient Mup1 delivery to the vacuole.

MeSH Terms
Endosomal Sorting Complexes Required for Transport/metabolism,physiology Membrane Proteins/genetics,metabolism Microscopy, Fluorescence Multivesicular Bodies/physiology Saccharomyces cerevisiae/genetics,metabolism,physiology Saccharomyces cerevisiae Proteins/genetics,metabolism Ubiquitin-Protein Ligase Complexes/metabolism Ubiquitination
Chemicals
Endosomal Sorting Complexes Required for Transport Membrane Proteins Saccharomyces cerevisiae Proteins Sna3 protein, S cerevisiae Ubiquitin-Protein Ligase Complexes RSP5 protein, S cerevisiae
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
MacDonald Chris
Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52246, USA.
Stringer Daniel K
Piper Robert C
References (54)
54 references, click to expand
  1. A concentric circle model of multivesicular body cargo sorting.
    EMBO Rep. 2007 Jul;8(7):644-50 PMID: 17603537
  2. Characterization of the WW domain of human yes-associated protein and its polyproline-containing ligands.
    J Biol Chem. 1997 Jul 4;272(27):17070-7 PMID: 9202023
  3. Multiple interactions drive adaptor-mediated recruitment of the ubiquitin ligase rsp5 to membrane proteins in vivo and in vitro.
    Mol Biol Cell. 2007 Jul;18(7):2429-40 PMID: 17429078
  4. Role of ubiquitylation in cellular membrane transport.
    Physiol Rev. 2006 Apr;86(2):669-707 PMID: 16601271
  5. Human immunodeficiency virus type 1 Nef protein targets CD4 to the multivesicular body pathway.
    J Virol. 2009 Jul;83(13):6578-90 PMID: 19403684
  6. Insights into ubiquitin transfer cascades from a structure of a UbcH5B approximately ubiquitin-HECT(NEDD4L) complex.
    Mol Cell. 2009 Dec 25;36(6):1095-102 PMID: 20064473
  7. Polyubiquitination by HECT E3s and the determinants of chain type specificity.
    Mol Cell Biol. 2009 Jun;29(12):3307-18 PMID: 19364824
  8. The membrane proteins, Spt23p and Mga2p, play distinct roles in the activation of Saccharomyces cerevisiae OLE1 gene expression. Fatty acid-mediated regulation of Mga2p activity is independent of its proteolytic processing into a soluble transcription activator.
    J Biol Chem. 2001 Nov 23;276(47):43548-56 PMID: 11557770
  9. An in vivo map of the yeast protein interactome.
    Science. 2008 Jun 13;320(5882):1465-70 PMID: 18467557
  10. Bsd2 binds the ubiquitin ligase Rsp5 and mediates the ubiquitination of transmembrane proteins.
    EMBO J. 2004 Mar 24;23(6):1279-88 PMID: 14988731
  11. Functional domains of the Rsp5 ubiquitin-protein ligase.
    Mol Cell Biol. 1999 Jan;19(1):342-52 PMID: 9858558
  12. Regulation of membrane protein transport by ubiquitin and ubiquitin-binding proteins.
    Annu Rev Cell Dev Biol. 2003;19:141-72 PMID: 14570567
  13. N-terminal ubiquitination: more protein substrates join in.
    Trends Cell Biol. 2004 Mar;14(3):103-6 PMID: 15055197
  14. Indirect immunofluorescence labeling in the yeast Saccharomyces cerevisiae.
    Cold Spring Harb Protoc. 2009 Nov;2009(11):pdb.prot5317 PMID: 20150056
  15. Two distinct pathways for targeting proteins from the cytoplasm to the vacuole/lysosome.
    J Cell Biol. 1997 Dec 29;139(7):1687-95 PMID: 9412464
  16. Characterization of multiple multivesicular body sorting determinants within Sna3: a role for the ubiquitin ligase Rsp5.
    Mol Biol Cell. 2007 Feb;18(2):707-20 PMID: 17182849
  17. E3 ubiquitin ligases as regulators of membrane protein trafficking and degradation.
    Traffic. 2005 Jun;6(6):429-41 PMID: 15882441
  18. Multivesicular body sorting: ubiquitin ligase Rsp5 is required for the modification and sorting of carboxypeptidase S.
    Mol Biol Cell. 2004 Feb;15(2):468-80 PMID: 14657247
  19. The ubiquitin code of yeast permease trafficking.
    Trends Cell Biol. 2010 Apr;20(4):196-204 PMID: 20138522
  20. Role of mammalian vacuolar protein-sorting proteins in endocytic trafficking of a non-ubiquitinated G protein-coupled receptor to lysosomes.
    J Biol Chem. 2004 May 21;279(21):22522-31 PMID: 15024011
  21. Direct binding to Rsp5p regulates ubiquitination-independent vacuolar transport of Sna3p.
    Mol Biol Cell. 2007 May;18(5):1781-9 PMID: 17332499
  22. How ubiquitin functions with ESCRTs.
    Traffic. 2011 Oct;12(10):1306-17 PMID: 21722280
  23. Identification of a PY motif in the epithelial Na channel subunits as a target sequence for mutations causing channel activation found in Liddle syndrome.
    EMBO J. 1996 May 15;15(10):2381-7 PMID: 8665845
  24. Breaking the chains: structure and function of the deubiquitinases.
    Nat Rev Mol Cell Biol. 2009 Aug;10(8):550-63 PMID: 19626045
  25. Direct binding to Rsp5 mediates ubiquitin-independent sorting of Sna3 via the multivesicular body pathway.
    Mol Biol Cell. 2007 Feb;18(2):697-706 PMID: 17182850
  26. Arrestin-mediated endocytosis of yeast plasma membrane transporters.
    Traffic. 2009 Dec;10(12):1856-67 PMID: 19912579
  27. Starvation induces vacuolar targeting and degradation of the tryptophan permease in yeast.
    J Cell Biol. 1999 Sep 20;146(6):1227-38 PMID: 10491387
  28. A novel sorting sequence in the beta2-adrenergic receptor switches recycling from default to the Hrs-dependent mechanism.
    J Biol Chem. 2007 Feb 2;282(5):3095-104 PMID: 17138565
  29. Transferrin-directed internalization and cycling of transferrin receptor 2.
    Traffic. 2009 Oct;10(10):1488-501 PMID: 19682329
  30. Ubiquitin-independent binding of Hrs mediates endosomal sorting of the interleukin-2 receptor beta-chain.
    J Cell Sci. 2008 May 15;121(Pt 10):1727-38 PMID: 18445679
  31. Characterization of a novel protein-binding module--the WW domain.
    FEBS Lett. 1995 Aug 1;369(1):67-71 PMID: 7641887
  32. Arrestin-like proteins mediate ubiquitination and endocytosis of the yeast metal transporter Smf1.
    EMBO Rep. 2008 Dec;9(12):1216-21 PMID: 18953286
  33. A single ubiquitin is sufficient for cargo protein entry into MVBs in the absence of ESCRT ubiquitination.
    J Cell Biol. 2011 Jan 24;192(2):229-42 PMID: 21242292
  34. Structure and function of a HECT domain ubiquitin-binding site.
    EMBO Rep. 2011 Apr;12(4):334-41 PMID: 21399621
  35. Sorting of proteins into multivesicular bodies: ubiquitin-dependent and -independent targeting.
    EMBO J. 2001 Sep 17;20(18):5176-86 PMID: 11566881
  36. The four-helix bundle of the neuronal target membrane SNARE complex is neither disordered in the middle nor uncoiled at the C-terminal region.
    J Biol Chem. 2002 Jul 5;277(27):24294-8 PMID: 11983696
  37. Arrestin-related ubiquitin-ligase adaptors regulate endocytosis and protein turnover at the cell surface.
    Cell. 2008 Nov 14;135(4):714-25 PMID: 18976803
  38. Targeting of Sna3p to the endosomal pathway depends on its interaction with Rsp5p and multivesicular body sorting on its ubiquitylation.
    Traffic. 2007 Sep;8(9):1280-96 PMID: 17645729
  39. The ESCRT machinery in endosomal sorting of ubiquitylated membrane proteins.
    Nature. 2009 Mar 26;458(7237):445-52 PMID: 19325624
  40. Ubiquitination of substrates by esterification.
    Traffic. 2012 Jan;13(1):19-24 PMID: 21883762
  41. The role of ubiquitination in lysosomal trafficking of δ-opioid receptors.
    Traffic. 2011 Feb;12(2):170-84 PMID: 21106040
  42. Ubiquitin is required for sorting to the vacuole of the yeast general amino acid permease, Gap1.
    J Biol Chem. 2001 Nov 23;276(47):43949-57 PMID: 11500494
  43. Genomic expression programs in the response of yeast cells to environmental changes.
    Mol Biol Cell. 2000 Dec;11(12):4241-57 PMID: 11102521
  44. Functional characterization of the S. cerevisiae genome by gene deletion and parallel analysis.
    Science. 1999 Aug 6;285(5429):901-6 PMID: 10436161
  45. Ubiquitin lys63 is involved in ubiquitination of a yeast plasma membrane protein.
    EMBO J. 1997 Oct 1;16(19):5847-54 PMID: 9312043
  46. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  47. Ubiquitination-independent trafficking of G protein-coupled receptors to lysosomes.
    J Biol Chem. 2002 Dec 27;277(52):50219-22 PMID: 12401797
  48. Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications.
    Yeast. 1998 Jan 30;14(2):115-32 PMID: 9483801
  49. Glucose-induced ubiquitylation and endocytosis of the yeast Jen1 transporter: role of lysine 63-linked ubiquitin chains.
    J Biol Chem. 2009 Jul 17;284(29):19228-36 PMID: 19433580
  50. Components of a ubiquitin ligase complex specify polyubiquitination and intracellular trafficking of the general amino acid permease.
    J Cell Biol. 2001 May 14;153(4):649-62 PMID: 11352928
  51. Physiological functions of the HECT family of ubiquitin ligases.
    Nat Rev Mol Cell Biol. 2009 Jun;10(6):398-409 PMID: 19436320
  52. Activation of a membrane-bound transcription factor by regulated ubiquitin/proteasome-dependent processing.
    Cell. 2000 Sep 1;102(5):577-86 PMID: 11007476
  53. The Vps4p AAA ATPase regulates membrane association of a Vps protein complex required for normal endosome function.
    EMBO J. 1998 Jun 1;17(11):2982-93 PMID: 9606181
  54. The PY-motif of Bul1 protein is essential for growth of Saccharomyces cerevisiae under various stress conditions.
    Gene. 1998 Dec 28;225(1-2):39-46 PMID: 9931424
Article Info
Journal
Traffic (Copenhagen, Denmark)
Abbr.
Traffic
ISSN
1600-0854
DOI
10.1111/j.1600-0854.2011.01326.x
Published
2012-04-00
Epub
2012-00-31
Pages
586-98
Language
English
Region
England
NLM ID
100939340
PMCID
PMC3336005
Subset
IM
Grants
NINDS NIH HHS · R21 NS064579-02 · United States
NINDS NIH HHS · R21 NS064579 · United States
NIGMS NIH HHS · R01GM58202 · United States
NIGMS NIH HHS · R01 GM058202 · United States
NIGMS NIH HHS · R01 GM058202-13 · United States
External Links
PubMed source DOI: 10.1111/j.1600-0854.2011.01326.x 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