Rescue of Munc18-1 and -2 double knockdown reveals the essential functions of interaction between Munc18 and closed syntaxin in PC12 cells

Han, Liping, Jiang, Tiandan, Han, Gayoung A., Malintan, Nancy T., Xie, Li., Wang, Li., Tse, Frederick W., Gaisano, Herbert Y., Collins, Brett M., Meunier, Frederic A. and Sugita, Shuzo (2009) Rescue of Munc18-1 and -2 double knockdown reveals the essential functions of interaction between Munc18 and closed syntaxin in PC12 cells. Molecular Biology of the Cell, 20 23: 4962-4975. doi:10.1091/mbc.E09-08-0712

Attached Files (Some files may be inaccessible until you login with your UQ eSpace credentials)
Name Description MIMEType Size Downloads
UQ192791_OA.pdf Full text (open access) application/pdf 3.47MB 0

Author Han, Liping
Jiang, Tiandan
Han, Gayoung A.
Malintan, Nancy T.
Xie, Li.
Wang, Li.
Tse, Frederick W.
Gaisano, Herbert Y.
Collins, Brett M.
Meunier, Frederic A.
Sugita, Shuzo
Title Rescue of Munc18-1 and -2 double knockdown reveals the essential functions of interaction between Munc18 and closed syntaxin in PC12 cells
Journal name Molecular Biology of the Cell   Check publisher's open access policy
ISSN 1059-1524
1939-4586
Publication date 2009-12-01
Sub-type Article (original research)
DOI 10.1091/mbc.E09-08-0712
Open Access Status File (Publisher version)
Volume 20
Issue 23
Start page 4962
End page 4975
Total pages 14
Editor Botstein, D.
Place of publication Bethesda, MD, United States
Publisher American Society for Cell Biology
Collection year 2010
Language eng
Subject C1
060108 Protein Trafficking
060110 Receptors and Membrane Biology
060105 Cell Neurochemistry
920111 Nervous System and Disorders
060112 Structural Biology (incl. Macromolecular Modelling)
Abstract Munc18-1 binds to syntaxin-1A via two distinct sites referred to as the "closed" conformation and N terminus binding. The latter has been shown to stimulate soluble N-ethylmaleimide-sensitive factor attachment protein receptor-mediated exocytosis, whereas the former is believed to be inhibitory or dispensable. To precisely define the contributions of each binding mode, we have engineered Munc18-1/-2 double knockdown neurosecretory cells and show that not only syntaxin-1A and -1B but also syntaxin-2 and -3 are significantly reduced as a result of Munc18-1 and -2 knockdown. Syntaxin-1 was mislocalized and the regulated secretion was abolished. We next examined the abilities of Munc18-1 mutants to rescue the defective phenotypes. Mutation (K46E/E59K) of Munc18-1 that selectively prevents binding to closed syntaxin-1 was unable to restore syntaxin-1 expression, localization, or secretion. In contrast, mutations (F115E/E132A) of Munc18-1 that selectively impair binding to the syntaxin-1 N terminus could still rescue the defective phenotypes. Our results indicate that Munc18-1 and -2 act in concert to support the expression of a broad range of syntaxins and to deliver syntaxin-1 to the plasma membrane. Our studies also indicate that the binding to the closed conformation of syntaxin is essential for Munc18-1 stimulatory action, whereas the binding to syntaxin N terminus plays a more limited role in neurosecretory cells.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 39 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 40 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Fri, 15 Jan 2010, 13:28:15 EST by Debra McMurtrie on behalf of Queensland Brain Institute