DEN1 deneddylates non-cullin proteins in vivo

The ubiquitin-like protein Nedd8/Rub1 covalently modifies and activates cullin ubiquitin ligases. However, the repertoire of Nedd8-modified proteins and the regulation of protein neddylation status are not clear. The cysteine protease DEN1/NEDP1 specifically processes the Nedd8 precursor and has been suggested to deconjugate Nedd8 from cullin proteins. By characterizing the Drosophila DEN1 protein and DEN1 null (DEN1null) mutants, we provide in vitro and in vivo evidence that DEN1, in addition to processing Nedd8, deneddylates many cellular proteins. Although purified DEN1 protein efficiently deneddylates the Nedd8-conjugated cullin proteins Cul1 and Cul3, neddylated Cul1 and Cul3 protein levels are not enhanced in DEN1null. Strikingly, many cellular proteins are highly neddylated in DEN1 mutants and are deneddylated by purified DEN1 protein. DEN1 deneddylation activity is distinct from that of the cullin-deneddylating CSN. Genetic analyses indicate that a balance between neddylation and deneddylation maintained by DEN1 is crucial for animal viability.

[1]  D. Xirodimas,et al.  Ribosomal proteins are targets for the NEDD8 pathway , 2008, EMBO reports.

[2]  M. Dasso,et al.  Modification in reverse: the SUMO proteases. , 2007, Trends in biochemical sciences.

[3]  M. Hochstrasser,et al.  Modification of proteins by ubiquitin and ubiquitin-like proteins. , 2006, Annual review of cell and developmental biology.

[4]  E. Yeh,et al.  Neddylation of a breast cancer-associated protein recruits a class III histone deacetylase that represses NFκB-dependent transcription , 2006, Nature Cell Biology.

[5]  C. Chien,et al.  Protection of cullin-RING E3 ligases by CSN-UBP12. , 2006, Trends in cell biology.

[6]  C. Chien,et al.  Neddylation and deneddylation regulate Cul1 and Cul3 protein accumulation , 2005, Nature Cell Biology.

[7]  F. Z. Watts,et al.  Nep1, a Schizosaccharomyces pombe deneddylating enzyme. , 2005, The Biochemical journal.

[8]  R. Hay,et al.  SUMO: a history of modification. , 2005, Molecular cell.

[9]  T. Toda,et al.  CSN facilitates Cullin–RING ubiquitin ligase function by counteracting autocatalytic adapter instability , 2005, Nature Cell Biology.

[10]  Keith D Wilkinson,et al.  Structure of a complex between Nedd8 and the Ulp/Senp protease family member Den1. , 2005, Journal of molecular biology.

[11]  D. Lane,et al.  Mdm2-Mediated NEDD8 Conjugation of p53 Inhibits Its Transcriptional Activity , 2004, Cell.

[12]  Erica S. Johnson,et al.  Protein modification by SUMO. , 2004, Annual review of biochemistry.

[13]  D. C. Dias,et al.  Nedd8 on cullin: building an expressway to protein destruction , 2004, Oncogene.

[14]  M. Bycroft,et al.  Structure of the Jab1/MPN domain and its implications for proteasome function. , 2003, Biochemistry.

[15]  Luming Yin,et al.  Identification and Characterization of DEN1, a Deneddylase of the ULP Family* , 2003, Journal of Biological Chemistry.

[16]  N. Wei,et al.  DEN1 Is a Dual Function Protease Capable of Processing the C Terminus of Nedd8 and Deconjugating Hyper-neddylated CUL1* , 2003, Journal of Biological Chemistry.

[17]  R. Hay,et al.  NEDP1, a Highly Conserved Cysteine Protease That deNEDDylates Cullins* , 2003, Journal of Biological Chemistry.

[18]  M. Mannervik,et al.  COP9 signalosome subunits 4 and 5 regulate multiple pleiotropic pathways in Drosophila melanogaster. , 2002, Development.

[19]  Cheng-Ting Chien,et al.  Distinct protein degradation mechanisms mediated by Cul1 and Cul3 controlling Ci stability in Drosophila eye development. , 2002, Genes & development.

[20]  J. B. Duffy,et al.  GAL4 system in drosophila: A fly geneticist's swiss army knife , 2002, Genesis.

[21]  L. Aravind,et al.  Role of Predicted Metalloprotease Motif of Jab1/Csn5 in Cleavage of Nedd8 from Cul1 , 2002, Science.

[22]  R. Deshaies,et al.  The fission yeast COP9/signalosome is involved in cullin modification by ubiquitin-related Ned8p , 2001, BMC Biochemistry.

[23]  A. Shevchenko,et al.  Promotion of NEDD8-CUL1 Conjugate Cleavage by COP9 Signalosome , 2001, Science.

[24]  Keiji Tanaka,et al.  Covalent modification of all members of human cullin family proteins by NEDD8 , 1999, Oncogene.

[25]  M. Tanouye,et al.  Drosophila homolog of APP-BP1 (dAPP-BP1) interacts antagonistically with APPL during Drosophila development , 2007, Cell Death and Differentiation.