Home LiteratureArticle Details
PMID: 16401725 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

An intramolecular t-SNARE complex functions in vivo without the syntaxin NH2-terminal regulatory domain.

The Journal of cell biology ·Vol. 172 ·No. 2 ·2006-01-16 ·Pages 295-307

Van Komen JS, Bai X, Scott BL, McNew JA

Abstract

Membrane fusion in the secretory pathway is mediated by SNAREs (located on the vesicle membrane [v-SNARE] and the target membrane [t-SNARE]). In all cases examined, t-SNARE function is provided as a three-helix bundle complex containing three approximately 70-amino acid SNARE motifs. One SNARE motif is provided by a syntaxin family member (the t-SNARE heavy chain), and the other two helices are contributed by additional t-SNARE light chains. The syntaxin family is the most conformationally dynamic group of SNAREs and appears to be the major focus of SNARE regulation. An NH2-terminal region of plasma membrane syntaxins has been assigned as a negative regulatory element in vitro. This region is absolutely required for syntaxin function in vivo. We now show that the required function of the NH2-terminal regulatory domain (NRD) of the yeast plasma membrane syntaxin, Sso1p, can be circumvented when t-SNARE complex formation is made intramolecular. Our results suggest that the NRD is required for efficient t-SNARE complex formation and does not recruit necessary scaffolding factors.

MeSH Terms
Amino Acid Sequence Membrane Fusion/physiology Multiprotein Complexes Point Mutation Protein Structure, Tertiary Qa-SNARE Proteins/chemistry,genetics,metabolism Qc-SNARE Proteins/genetics,metabolism Recombinant Fusion Proteins/genetics,metabolism SNARE Proteins/chemistry,genetics,metabolism Saccharomyces cerevisiae/cytology,genetics,metabolism Saccharomyces cerevisiae Proteins/chemistry,genetics,metabolism
Chemicals
Multiprotein Complexes Qa-SNARE Proteins Qc-SNARE Proteins Recombinant Fusion Proteins SEC9 protein, S cerevisiae SNARE Proteins SSO1 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Van Komen Jeffrey S
Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77251, USA.
Bai Xiaoyang
Scott Brenton L
McNew James A
References (40)
40 references, click to expand
  1. Rapid and efficient fusion of phospholipid vesicles by the alpha-helical core of a SNARE complex in the absence of an N-terminal regulatory domain.
    Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12565-70 PMID: 10535962
  2. New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae.
    Yeast. 1994 Dec;10(13):1793-808 PMID: 7747518
  3. Structural analysis of the neuronal SNARE protein syntaxin-1A.
    Biochemistry. 2000 Jul 25;39(29):8470-9 PMID: 10913252
  4. Compartmental specificity of cellular membrane fusion encoded in SNARE proteins.
    Nature. 2000 Sep 14;407(6801):153-9 PMID: 11001046
  5. Functional architecture of an intracellular membrane t-SNARE.
    Nature. 2000 Sep 14;407(6801):198-202 PMID: 11001059
  6. The N-ethylmaleimide-sensitive fusion protein and alpha-SNAP induce a conformational change in syntaxin.
    J Biol Chem. 1995 Jul 14;270(28):16955-61 PMID: 7622514
  7. A structural change occurs upon binding of syntaxin to SNAP-25.
    J Biol Chem. 1997 Feb 14;272(7):4582-90 PMID: 9020186
  8. Formation of a yeast SNARE complex is accompanied by significant structural changes.
    FEBS Lett. 1997 Sep 22;415(1):49-55 PMID: 9326367
  9. Structural changes are associated with soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor complex formation.
    J Biol Chem. 1997 Oct 31;272(44):28036-41 PMID: 9346956
  10. Yeast spore germination: a requirement for Ras protein activity during re-entry into the cell cycle.
    EMBO J. 1997 Oct 15;16(20):6171-81 PMID: 9321396
  11. SNAREpins: minimal machinery for membrane fusion.
    Cell. 1998 Mar 20;92(6):759-72 PMID: 9529252
  12. Regulation of SNARE complex assembly by an N-terminal domain of the t-SNARE Sso1p.
    Nat Struct Biol. 1998 Sep;5(9):793-802 PMID: 9731774
  13. Three-dimensional structure of an evolutionarily conserved N-terminal domain of syntaxin 1A.
    Cell. 1998 Sep 18;94(6):841-9 PMID: 9753330
  14. Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 A resolution.
    Nature. 1998 Sep 24;395(6700):347-53 PMID: 9759724
  15. Folding intermediates of SNARE complex assembly.
    Nat Struct Biol. 1999 Feb;6(2):117-23 PMID: 10048921
  16. Yeast homologues of tomosyn and lethal giant larvae function in exocytosis and are associated with the plasma membrane SNARE, Sec9.
    J Cell Biol. 1999 Jul 12;146(1):125-40 PMID: 10402465
  17. Interactions within the yeast t-SNARE Sso1p that control SNARE complex assembly.
    Nat Struct Biol. 2000 Oct;7(10):894-902 PMID: 11017200
  18. Testing the 3Q:1R "rule": mutational analysis of the ionic "zero" layer in the yeast exocytic SNARE complex reveals no requirement for arginine.
    Mol Biol Cell. 2000 Nov;11(11):3849-58 PMID: 11071911
  19. SNARE-mediated membrane fusion.
    Nat Rev Mol Cell Biol. 2001 Feb;2(2):98-106 PMID: 11252968
  20. Homo- and heterooligomeric SNARE complexes studied by site-directed spin labeling.
    J Biol Chem. 2001 Apr 20;276(16):13169-77 PMID: 11278719
  21. The syntaxins.
    Genome Biol. 2001;2(11):REVIEWS3012 PMID: 11737951
  22. Characterization of temperature-sensitive mutations in the yeast syntaxin 1 homologues Sso1p and Sso2p, and evidence of a distinct function for Sso1p in sporulation.
    J Cell Sci. 2002 Jan 15;115(Pt 2):409-20 PMID: 11839791
  23. Conformational regulation of SNARE assembly and disassembly in vivo.
    J Biol Chem. 2002 Mar 15;277(11):9375-81 PMID: 11777922
  24. 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
  25. Membrane topologies of neuronal SNARE folding intermediates.
    Biochemistry. 2002 Sep 10;41(36):10928-33 PMID: 12206663
  26. Parallel phenotypic analysis of sporulation and postgermination growth in Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 2002 Nov 26;99(24):15530-5 PMID: 12432101
  27. Hrs regulates early endosome fusion by inhibiting formation of an endosomal SNARE complex.
    J Cell Biol. 2003 Jul 7;162(1):125-37 PMID: 12847087
  28. SNARE protein structure and function.
    Annu Rev Cell Dev Biol. 2003;19:493-517 PMID: 14570579
  29. Liposome fusion assay to monitor intracellular membrane fusion machines.
    Methods Enzymol. 2003;372:274-300 PMID: 14610819
  30. Single-molecule fluorescence resonance energy transfer reveals a dynamic equilibrium between closed and open conformations of syntaxin 1.
    Proc Natl Acad Sci U S A. 2003 Dec 23;100(26):15516-21 PMID: 14668446
  31. Constitutive versus regulated SNARE assembly: a structural basis.
    EMBO J. 2004 Feb 25;23(4):681-9 PMID: 14765122
  32. Sec1p directly stimulates SNARE-mediated membrane fusion in vitro.
    J Cell Biol. 2004 Oct 11;167(1):75-85 PMID: 15466482
  33. Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway.
    Cell. 1980 Aug;21(1):205-15 PMID: 6996832
  34. 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
  35. High resolution analysis of functional determinants on human tissue-type plasminogen activator.
    J Biol Chem. 1991 Mar 15;266(8):5191-201 PMID: 1900516
  36. Rational scanning mutagenesis of a protein kinase identifies functional regions involved in catalysis and substrate interactions.
    J Biol Chem. 1991 May 15;266(14):8923-31 PMID: 2026604
  37. Yeast syntaxins Sso1p and Sso2p belong to a family of related membrane proteins that function in vesicular transport.
    EMBO J. 1993 Nov;12(11):4095-104 PMID: 8223426
  38. Protein-protein interactions contributing to the specificity of intracellular vesicular trafficking.
    Science. 1994 Feb 25;263(5150):1146-9 PMID: 8108733
  39. A rab protein is required for the assembly of SNARE complexes in the docking of transport vesicles.
    Cell. 1994 Sep 23;78(6):937-48 PMID: 7923363
  40. Three-dimensional structure of the neuronal-Sec1-syntaxin 1a complex.
    Nature. 2000 Mar 23;404(6776):355-62 PMID: 10746715
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200507138
Published
2006-01-16
Epub
2006-00-09
Pages
295-307
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2063558
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.200507138 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