Mono-ubiquitination drives nuclear export of the human DCN1-like protein hDCNL Journal Article


Authors: Wu, K.; Yan, H.; Fang, L.; Wang, X.; Pfleger, C.; Jiang, X.; Huang, L.; Pan, Z. Q.
Article Title: Mono-ubiquitination drives nuclear export of the human DCN1-like protein hDCNL
Abstract: Conjugation of Nedd8 to a cullin protein, termed neddylation, is an evolutionarily conserved process that functions to activate the cullin-RING family E3 ubiquitin ligases, leading to increased proteasomal degradation of a wide range of substrate proteins. Recent emerging evidence demonstrates that cellular neddylation requires the action of Dcn1, which, in humans, consists of five homologues designated as hDCNL1-5. Here we revealed a previously unknown mechanism that regulates hDCNL1. In cultured mammalian cells ectopically expressed hDCNL1 was mono-ubiquitinated predominantly at K143, K149, and K171. Using a classical chromatographic purification strategy, we identified Nedd4-1 as an E3 ligase that can catalyze mono-ubiquitination of hDCNL1 in a reconstituted ubiquitination system. In addition, the hDCNL1 N-terminal ubiquitin-binding domain is necessary and sufficient to mediate mono-ubiquitination. Finally, fluorescence microscopic and subcellular fractionation analyses revealed a role for mono-ubiquitination in driving nuclear export of hDCNL1. Taken together, these results suggest a mono-ubiquitination-mediated mechanism that governs nuclear-cytoplasmic trafficking of hDCNL1, thereby regulating hDCNL1-dependent activation of the cullin-RING E3 ubiquitin ligases in selected cellular compartments. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.
Keywords: controlled study; protein expression; unclassified drug; human cell; proto-oncogene proteins; proteins; mammalia; embryo; hela cells; nucleocytoplasmic transport; ubiquitination; amino terminal sequence; cell culture; protein transport; cytoplasm; active transport, cell nucleus; binding protein; binding site; cell fractionation; protein structure, tertiary; cell nucleus; cell transport; mammals; ubiquitin protein ligase nedd4; endosomal sorting complexes required for transport; ubiquitin-protein ligases; enzymes; lysine; mammal cell; e3 ligase; nuclear export; hek293 cells; n-terminals; proteasomal degradation; cellular compartments; chromatographic purification; e3 ubiquitin ligases; mammalian cells; subcellular fractionation; substrate proteins; chromatographic analysis; dcn1 like protein
Journal Title: Journal of Behavioral Medicine
Volume: 286
Issue: 39
ISSN: 0160-7715
Publisher: Springer  
Date Published: 2011-09-30
Start Page: 34060
End Page: 34070
Language: English
DOI: 10.1074/jbc.M111.273045
PROVIDER: scopus
PMCID: PMC3190805
PUBMED: 21813641
DOI/URL:
Notes: --- - "Export Date: 2 November 2011" - "CODEN: JBCHA" - "Source: Scopus"
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MSK Authors
  1. Xinjiang Wang
    6 Wang
  2. Xuejun Jiang
    93 Jiang