The ubiquitin ligase itch is auto-ubiquitylated in vivo and in vitro but is protected from degradation by interacting with the deubiquitylating enzyme FAM/USP9X

Mouchantaf, Rania, Azakir, Bilal A., McPherson, Peter S., Millard, Susan M., Wood, Stephen A. and Angers, Annie (2006) The ubiquitin ligase itch is auto-ubiquitylated in vivo and in vitro but is protected from degradation by interacting with the deubiquitylating enzyme FAM/USP9X. Journal of Biological Chemistry, 281 50: 38738-38747. doi:10.1074/jbc.M605959200

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Author Mouchantaf, Rania
Azakir, Bilal A.
McPherson, Peter S.
Millard, Susan M.
Wood, Stephen A.
Angers, Annie
Title The ubiquitin ligase itch is auto-ubiquitylated in vivo and in vitro but is protected from degradation by interacting with the deubiquitylating enzyme FAM/USP9X
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
Publication date 2006-12-01
Sub-type Article (original research)
DOI 10.1074/jbc.M605959200
Open Access Status File (Publisher version)
Volume 281
Issue 50
Start page 38738
End page 38747
Total pages 10
Place of publication Bethesda, MD, United States
Publisher American society for Biochemistry and Molecular Biology
Language eng
Formatted abstract
Itch is a ubiquitin ligase that has been implicated in the regulation of a number of cellular processes. We previously have identified Itch as a binding partner for the endocytic protein Endophilin and found it to be localized to endosomes. Using affinity purification coupled to mass spectrometry, we have now identified the ubiquitin-protease FAM/USP9X as a binding partner of Itch. The association between Itch and FAM/USP9X was confirmed in vitro by glutathione S-transferase pulldown and in vivo through coimmunoprecipation. Itch and FAM partially colocalize in COS-7 cells at the trans-Golgi network and in peripheral vesicles. We mapped the FAM-binding domain on Itch to the WW domains, a region known to be involved in substrate recognition. However, transient overexpression of FAM/USP9X resulted in the deubiquitylation of Itch. Moreover, we show that Itch auto-ubiquitylation leads to its degradation in the proteasome. By examining the amounts of Itch and FAM in various cell lines and rat tissues, a positive correlation was found in the expression of both proteins. This observation suggests that the levels of FAM expression could have an influence on Itch in cells. Experimental decrease in FAM levels by RNA interference leads to a significant reduction in intracellular levels of endogenous Itch, which can be prevented by treatment with the proteasome inhibitor lactacystin. Accordingly, overexpression of FAM/USP9X resulted in a marked increase in endogenous Itch levels. These results demonstrate an intriguing interplay between a ubiquitin ligase and a ubiquitin protease, based on direct interaction between the two proteins. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.
Keyword Facets Deubiquitinating Enzyme
Fat-Facets
E3 Ligase
Embryo Development
Down-Regulation
Jnk Activation
Liquid Facets
Beta-Catenin
Protein
Domain
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collections: UQ Centre for Clinical Research Publications
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