Identification of the activating and conjugating enzymes of the NEDD8 conjugation pathway

J Biol Chem. 1999 Apr 23;274(17):12036-42. doi: 10.1074/jbc.274.17.12036.


NEDD8 is a ubiquitin-like molecule that can be covalently conjugated to a limited number of cellular proteins, such as Cdc53/cullin. We have previously reported that the C terminus of NEDD8 is efficiently processed to expose Gly-76, which is required for conjugation to target proteins. A combination of data base searches and polymerase chain reaction cloning was used to identify a cDNA encoding human UBA3, which is 38% identical to the yeast homologue, 22% identical to human UBA2, and 19% identical to the C-terminal region of human UBE1. The human UBA3 gene is located on chromosome 3p13 and gave rise to a 2.2-kilobase pair transcript that was detected in all tissues. Human UBA3 could be precipitated with glutathione S-transferase (GST)-NEDD8, but not with GST-ubiquitin or GST-sentrin-1. Moreover, human UBA3 could form a beta-mercaptoethanol-sensitive conjugate with NEDD8 in the presence of APP-BP1, a protein with sequence homology to the N-terminal half of ubiquitin-activating enzyme. We have also cloned human UBC12 and demonstrated that it could form a thiol ester linkage with NEDD8 in the presence of the activating enzyme complex. Identification of the activating and conjugating enzymes of the NEDD8 conjugation pathway should allow for a more detailed study of the role of NEDD8 modification in health and disease.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Chromosome Mapping
  • Chromosomes, Human, Pair 3
  • Cloning, Molecular
  • DNA, Complementary
  • Humans
  • Ligases / metabolism*
  • Mice
  • Molecular Sequence Data
  • NEDD8 Protein
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae Proteins*
  • Sequence Homology, Amino Acid
  • Ubiquitin-Activating Enzymes
  • Ubiquitins / genetics
  • Ubiquitins / metabolism*


  • DNA, Complementary
  • NEDD8 Protein
  • NEDD8 protein, human
  • Nedd8 protein, mouse
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Saccharomyces cerevisiae Proteins
  • UBA3 protein, S cerevisiae
  • Ube1c protein, mouse
  • Ubiquitins
  • Ligases
  • Ubiquitin-Activating Enzymes
  • NAE protein, human

Associated data

  • GENBANK/AF046024