Molecular cloning of the cDNA and chromosome localization of the gene for human ubiquitin-conjugating enzyme 9.
We report a novel human gene whose product specifically associates with the negative regulatory domain of the Wilms' tumor gene product (WT1) in a yeast two-hybrid screen and with WT1 in immunoprecipitation and glutathione S-transferase (GST) capture assays. The gene encodes a 17-kDa protein that has 56% amino acid sequence identity with yeast ubiquitin-conjugating enzyme (yUBC) 9, a protein required for cell cycle progression in yeast, and significant identity with other subfamilies of ubiquitin-conjugating enzymes. The human gene fully complements yeast that have a temperature-sensitive yUBC9 gene mutation to fully restore normal growth, indicating that we have cloned a functionally conserved human (h) homolog of yUBC9. Transcripts of hUBC9 of 4.4 kilobases (kb), 2.8 kb, and 1.3 kb were found in all human tissues tested. A single copy of the hUBC9 gene was found and localized to human chromosome 16p13.3. We conclude that hUBC9 retains striking structural and functional conservation with yUBC9 and suggest a possible link of the ubiquitin/proteosome proteolytic pathway and the WT1 transcriptional repressor system.
Wang
Z Y
ZY
Department of Medicine, Beth Israel Hospital and the Harvard Medical School, Boston, Massachusetts 02215, USA.
Qiu
Q Q
QQ
Seufert
W
W
Taguchi
T
T
Testa
J R
JR
Whitmore
S A
SA
Callen
D F
DF
Welsh
D
D
Shenk
T
T
Deuel
T F
TF
D000595:Amino Acid Sequence; D002874:Chromosome Mapping; D002885:Chromosomes, Human, Pair 16; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D008025:Ligases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes; D025721:WT1 Proteins; D000098843:Ubiquitin-Conjugating Enzyme UBC9
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D014157:Transcription Factors; D025721:WT1 Proteins; D044763:Ubiquitin-Conjugating Enzymes; D008025:Ligases; D000098843:Ubiquitin-Conjugating Enzyme UBC9