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PMID: 11782429 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

LNX functions as a RING type E3 ubiquitin ligase that targets the cell fate determinant Numb for ubiquitin-dependent degradation.

The EMBO journal ·Vol. 21 ·No. 1-2 ·2002-01-15 ·Pages 93-102

Nie J, McGill MA, Dermer M, Dho SE, Wolting CD, McGlade CJ

Abstract

LNX is a RING finger and PDZ domain containing protein that interacts with the cell fate determinant Numb. To investigate the function of LNX, we tested its RING finger domain for ubiquitin ligase activity. The isolated RING finger domain was able to function as an E2-dependent, E3 ubiquitin ligase in vitro and mutation of a conserved cysteine residue within the RING domain abolished its activity, indicating that LNX is the first described PDZ domain-containing member of the E3 ubiquitin ligase family. We have identified Numb as a substrate of LNX E3 activity in vitro and in vivo. In addition to the RING finger, a region of LNX, including the Numb PTB domain-binding site and the first PDZ domain, is required for Numb ubiquitylation. Expression of wild-type but not mutant LNX causes proteasome-dependent degradation of Numb and can enhance Notch signalling. These results suggest that the levels of mammalian Numb protein and therefore, by extension, the processes of asymmetric cell division and cell fate determination may be regulated by ubiquitin-dependent proteolysis.

MeSH Terms
Animals Binding Sites CHO Cells Carrier Proteins/chemistry,genetics,metabolism Cell Line Cricetinae Dogs Humans Ligases/genetics,metabolism Membrane Proteins/genetics,metabolism Mice Mutagenesis, Site-Directed Nerve Tissue Proteins/genetics,metabolism Protein Structure, Tertiary Recombinant Fusion Proteins/chemistry,genetics,metabolism Substrate Specificity Ubiquitin/metabolism Ubiquitin-Protein Ligases
Chemicals
Carrier Proteins Membrane Proteins Nerve Tissue Proteins Numb protein, mouse Recombinant Fusion Proteins Ubiquitin LNX1 protein, human Lnx1 protein, mouse Ubiquitin-Protein Ligases Ligases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Nie Jing
The Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children and Department of Medical Biophysics, University of Toronto, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada.
McGill Melanie A
Dermer Matt
Dho Sascha E
Wolting Cheryl D
McGlade C Jane
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/21.1.93
Published
2002-01-15
Pages
93-102
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125803
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/21.1.93 PMC Full Text View journal details
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