Hrs interacts with SNAP-25 and regulates Ca2+-dependent exocytosis

Jeffrey Kwong, Francine L. Roudabush, P. Hutton Moore, Michael Montague, William Oldham, Yankun Li, Lih Shen Chin, Lian Li

Research output: Contribution to journalArticle

Abstract

Synaptosome-associated protein of 25 kDa (SNAP-25) is a neuronal membrane protein essential for synaptic vesicle exocytosis. To investigate the mechanisms by which SNAP-25 mediates neurosecretion, we performed a search for proteins that interact with SNAP-25 using a yeast two-hybrid screen. Here, we report the isolation and characterization of a SNAP-25-interacting protein that is the rat homologue of mouse hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs). Hrs specifically interacts with SNAP-25, but not SNAP-23/syndet. The association of Hrs and SNAP-25 is mediated via coiled-coil interactions. Using an Hrs-specific antibody, we have shown that Hrs is highly enriched in brain, where it codistributes with SNAP-25 in most brain regions. Subcellular fractionation studies demonstrate that in brain, Hrs exists in both cytosolic and membrane-associated pools. Studies using indirect immunofluorescence and confocal microscopy reveal that, in addition to early endosomes, Hrs is also localized to large dense-core secretory granules and synaptic-like microvesicles in nerve growth factor-differentiated PC12 cells. Moreover, overexpression of Hrs in PC12 cells inhibits Ca2+-dependent exocytosis. These results suggest that Hrs is involved in regulation of neurosecretion through interaction with SNAP-25.

Original languageEnglish (US)
Pages (from-to)2273-2284
Number of pages12
JournalJournal of cell science
Volume113
Issue number12
StatePublished - Jul 13 2000
Externally publishedYes

Keywords

  • Growth factor
  • Hrs
  • SNAP-25
  • SNARE
  • Vesicular transport
  • Yeast two-hybrid screen

ASJC Scopus subject areas

  • Cell Biology

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  • Cite this

    Kwong, J., Roudabush, F. L., Moore, P. H., Montague, M., Oldham, W., Li, Y., Chin, L. S., & Li, L. (2000). Hrs interacts with SNAP-25 and regulates Ca2+-dependent exocytosis. Journal of cell science, 113(12), 2273-2284.