Jab1, a novel protease-activated receptor-2 (PAR-2)-interacting protein, is involved in PAR-2-induced activation of activator protein-1

Weibo Luo, Yingfei Wang, Theodor Hanck, Rolf Stricker, Georg Reiser

Research output: Contribution to journalArticle

Abstract

Protease-activated receptor-2 (PAR-2), a G protein-coupled receptor for trypsin and tryptase, exerts important physiological and pathological functions in multiple systems. However, unlike PAR-1, the PAR-2-mediated intracellular signal transductions are hardly known. Here, using yeast two-hybrid screening with a human brain cDNA library, we identified an interacting partner of human PAR-2, the Jun activation domain-binding protein 1 (Jab1). The interaction was confirmed by glutathione S-transferase pull-down assays in vitro, and by co-immunoprecipitation assays in vivo. Jab1 was also shown to be colocalized with PAR-2 in both transfected HEK293 cells and in normal primary human astrocytes by double immunofluorescence staining. Further experiments demonstrated that multiple intracellular domains of PAR-2 are required for the interaction with Jab1. We then showed that agonist stimulation of PAR-2 disrupted the interaction, which could be prevented by the inhibitor of receptor endocytosis phenylarsine oxide, but not by the lysosomal protease inhibitor ZPAD. Importantly, we found that activation of PAR-2 induced the redistribution of Jab1 from the plasma membrane to the cytosol, but did not influence expression of Jab1. Furthermore, Jab1 mediated PAR-2-induced c-Jun activation, which was followed by increased activation of activator protein-1. Loss-of-function studies, using Jab1 small interfering RNA, demonstrated that Jab1 knockdown blocked PAR-2-induced activator protein-1 activation. Taken together, our data demonstrate that Jab1 is an important effector that mediates a novel signal transduction pathway for PAR-2-dependent gene expression.

Original languageEnglish (US)
Pages (from-to)7927-7936
Number of pages10
JournalJournal of Biological Chemistry
Volume281
Issue number12
DOIs
StatePublished - Mar 24 2006
Externally publishedYes

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Receptor-Interacting Protein Serine-Threonine Kinases
PAR-2 Receptor
Transcription Factor AP-1
Carrier Proteins
Chemical activation
Proteins
Signal transduction
Signal Transduction
Assays
Tryptases
HEK293 Cells
G-Protein-Coupled Receptors
Endocytosis
Protease Inhibitors
Glutathione Transferase
Gene Library
Immunoprecipitation
Astrocytes
Cytosol
Trypsin

ASJC Scopus subject areas

  • Biochemistry

Cite this

Jab1, a novel protease-activated receptor-2 (PAR-2)-interacting protein, is involved in PAR-2-induced activation of activator protein-1. / Luo, Weibo; Wang, Yingfei; Hanck, Theodor; Stricker, Rolf; Reiser, Georg.

In: Journal of Biological Chemistry, Vol. 281, No. 12, 24.03.2006, p. 7927-7936.

Research output: Contribution to journalArticle

Luo, Weibo ; Wang, Yingfei ; Hanck, Theodor ; Stricker, Rolf ; Reiser, Georg. / Jab1, a novel protease-activated receptor-2 (PAR-2)-interacting protein, is involved in PAR-2-induced activation of activator protein-1. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 12. pp. 7927-7936.
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