Abstract
The ubiquitin-protein conjugation system is involved in a variety of eukaryotic cell functions, including the degradation of abnormal and short-lived proteins, chromatin structure, cell cycle progression, and DNA repair. The ubiquitination of target proteins is catalyzed by a ubiquitin-activating enzyme (E1) and ubiquitin-conjugating enzymes (E2s) and in some cases also requires auxiliary substrate recognition proteins (E3s). Multiple E2s have been found, and these likely possess specificity for different classes of target proteins. Here we report the cloning and characterization of a gene (ubc-2) encoding a ubiquitin-conjugating enzyme which is involved in the selective degradation of abnormal and short-lived proteins in the nematode Caenorhabditis elegans. The nematode ubc-2 gene encodes a 16.7-kDa protein with striking amino acid sequence similarity to Saccharomyces cerevisiae UBC4 and UBC5 and Drosophila UbcD1. When driven by the UBC4 promoter, ubc-2 can functionally substitute for UBC4 in yeast cells; it rescues the slow-growth phenotype of ubc4 ubc5 mutants at normal temperature and restores their ability to grow at elevated temperatures. Western blots (immunoblots) of ubc4 ubc5 yeast cells transformed with ubc-2 reveal a protein of the expected size, which cross-reacts with anti-Drosophila UbcD1 antibody. C. elegans ubc-2 is constitutively expressed at all life cycle stages and, unlike yeast UBC4 and UBC5, is not induced by heat shock. Both trans and cis splicing are involved in the maturation of the ubc-2 transcript. These data suggest that yeast UBC4 and UBC5, Drosophila UbcD1, and C. elegans ubc-2 define a highly conserved gene family which plays fundamental roles in all eukaryotic cells.
MeSH Terms
Amino Acid Sequence
Animals
Base Sequence
Caenorhabditis elegans/enzymology,genetics,growth & development
Caenorhabditis elegans Proteins
Cloning, Molecular
Drosophila/genetics
Genes, Helminth/genetics
Genetic Complementation Test
Helminth Proteins/metabolism
Hot Temperature
Ligases/genetics,metabolism
Molecular Sequence Data
Morphogenesis
RNA Precursors/metabolism
RNA Splicing
Saccharomyces cerevisiae/genetics
Saccharomyces cerevisiae Proteins
Sequence Homology, Amino Acid
Ubiquitin-Conjugating Enzymes
Chemicals
Caenorhabditis elegans Proteins
Helminth Proteins
RNA Precursors
Saccharomyces cerevisiae Proteins
UBC5 protein, S cerevisiae
Ubiquitin-Conjugating Enzymes
let-70 protein, C elegans
ubiquitin-conjugating enzyme UBC4
Ligases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Zhen M
Department of Biochemistry, University of British Columbia, Vancouver, Canada.
Heinlein R
Jones D
Jentsch S
Candido E P
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