Ubiquitylation of terminal deoxynucleotidyltransferase inhibits its activity.
Terminal deoxynucleotidyltransferase (TdT), which template-independently synthesizes DNA during V(D)J recombination in lymphoid cells, is ubiquitylated by a BPOZ-2/Cul3 complex, as the ubiquitin ligase, and then degraded by the 26 S proteasome. We show here that TdT is ubiquitylated by the Cul3-based ubiquitylation system in vitro. Because TdT could also be ubiquitylated in the absence of Cul/BPOZ-2, we determined that it could also be directly ubiquitylated by the E2 proteins UbcH5a/b/c and UbcH6, E3-independently. Furthermore, the ubiquitylated TdT inhibited its nucleotidyltransferase activity.
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda, Chiba, Japan. maezws@rs.noda.tus.ac.jp
So
S
Maezawa
Rie
R
Fukushima
Toyofumi
T
Matsushita
Tomoyoshi
T
Kato
Yoshiki
Y
Takagaki
Yoshihiro
Y
Nishiyama
Sachiko
S
Ando
Takuro
T
Matsumoto
Kousuke
K
Kouda
Takahide
T
Hayano
Masahiro
M
Suzuki
Kotaro
K
Koiwai
Osamu
O
Koiwai
D000818:Animals; D002417:Cattle; D044842:Cullin Proteins / Q000235:genetics / Q000378:metabolism; D004253:DNA Nucleotidylexotransferase / Q000235:genetics / Q000378:metabolism*; D025461:Feedback, Physiological; D005786:Gene Expression Regulation; D015723:Gene Library; D006367:HeLa Cells; D006801:Humans; D008099:Liver / Q000166:cytology / Q000378:metabolism; D010957:Plasmids; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D059748:Proteolysis; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology / Q000378:metabolism; D014162:Transfection; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination / Q000235:genetics*
C402243:ABTB1 protein, human; C503645:CUL3 protein, human; D044842:Cullin Proteins; D012097:Repressor Proteins; C097308:UBE2D1 protein, human; C097309:UBE2D2 protein, human; C097310:UBE2D3 protein, human; D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases; D004253:DNA Nucleotidylexotransferase; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease