The E3 ubiquitin ligase Itch regulates sorting nexin 9 through an unconventional substrate recognition domain.
The level of intracellular proteins is mainly regulated through modifications by ubiquitin ligases that target them for degradation. Members of the NEDD4 family of E3 ubiquitin ligases, such as Itch (atrophin-1 interacting protein 4), possess up to four WW domains for specific association with PY motif-containing substrates. We have identified sorting nexin 9 (SNX9), a protein involved in endocytic processes, as a new substrate of Itch. Itch ubiquitylates SNX9 and regulates intracellular SNX9 levels. Using truncated proteins, we found that the interaction with SNX9 is mediated by the proline-rich domain (PRD) of Itch, a domain distinct from the conventional WW recognition domain, and the SH3 domain of SNX9. Interaction with the PRD of Itch is essential for SNX9 ubiquitylation and degradation. Furthermore, this effect is specific for Itch, as NEDD4, a related PRD-containing E3 ligase, does not bind SNX9. SNX18, a second member of the SNX family containing an SH3 domain, was also found to bind to Itch. Our results indicate that the pool of substrates of NEDD4 family E3 ubiquitin ligases extends beyond proteins containing PY motifs.
Baumann
Claudia
C
Ludwig Institute for Cancer Research Ltd, University of Lausanne, Lausanne, Switzerland. Claudia.Baumann@licr.unil.ch
Lindholm
Cecilia K
CK
Rimoldi
Donata
D
Lévy
Frédéric
F
D002460:Cell Line; D006801:Humans; D055232:Proline-Rich Protein Domains; D012097:Repressor Proteins / Q000737:chemistry* / Q000378:metabolism*; D058305:Sorting Nexins; D013379:Substrate Specificity; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry* / Q000378:metabolism*; D033921:Vesicular Transport Proteins / Q000737:chemistry* / Q000378:metabolism*; D018909:src Homology Domains
D012097:Repressor Proteins; C400533:SNX9 protein, human; D058305:Sorting Nexins; D033921:Vesicular Transport Proteins; C432966:ITCH protein, human; D044767:Ubiquitin-Protein Ligases