[The identification and cloning of human ubiquitin binding enzyme cDNA].

OBJECTIVE

To identify and clone the gene encoding human ubiquitin binding enzyme and study its expression spectrum.

METHODS

According to the sequence of human EST, which is highly homologous to the mouse ubiquitin conjugating enzyme (E2), primers used for library screening were synthesized to screen the human fetal brain cDNA library. The gene was analyzed by making use of bioinformatics and its expression spectrum was studied by using multiple-tissue Northern blot.

RESULTS

Two cDNA clones encoding human ubiquitin conjugating enzyme were isolated and identified. Both containing the ubiquitin conjugating domain, they were 88% identical in amino acid sequences and found to be isoforms of each other with only an exon excised from the short sequence. They belonged to a highly conserved, and widely expressed E2 enzyme family. Northern blot showed that they were expressed exclusively in heart, placenta, and pancreas while no transcripts could be detected in brain, lung, liver, skeletal muscle, or kidney.

CONCLUSIONS

The gene encoding human ubiquitin binding enzyme is expressed under both temporal and spatial control. As a key enzyme in the degradation of proteins, ubiquitin conjugating enzymes play a central role in the expression regulation on the level of post-translation.