PMID- 10066434
OWN - NLM
STAT- MEDLINE
DCOM- 19990413
LR  - 20161124
IS  - 0006-291X (Print)
IS  - 0006-291X (Linking)
VI  - 256
IP  - 1
DP  - 1999 Mar 5
TI  - In vivo and in vitro recruitment of an IkappaBalpha-ubiquitin ligase to
      IkappaBalpha phosphorylated by IKK, leading to ubiquitination.
PG  - 121-6
AB  - Activation of the transcriptional factor NF-kappaB is triggered by
      signal-dependent degradation of its inhibitor protein IkappaB through the
      ubiquitin (Ub)-proteasome pathway. We found here that a phosphorylated
      IkappaBalpha immunoprecipitated (IP-pIkappaBalpha) from the crude extract of HeLa
      cells which had been treated with tumor necrosis factor-alpha (TNFalpha) caused a
      dramatic ubiquitination of itself, termed autoubiquitination, when incubated with
      ATP, Ub, and E1-activating and E2-conjugating enzymes. IP-pIkappaBalpha also
      catalyzed ubiquitination of an in vitro synthesized 35S-IkappaBalpha previously
      phosphorylated by IkappaB-kinase (IKK) which is referred to as
      transubiquitination. No appreciable activity of auto- and transubiquitination was
      observed in an unphosphorylated IP-IkappaBalpha. Moreover, the putative
      IkappaBalpha-Ub ligase (IkappaBalpha-E3) present in HeLa cell cytosol associated 
      in vitro with an IKK-phosphorylated recombinant IkappaBalpha, a process
      independent of NF-kappaB binding to IkappaBalpha or TNFalpha stimulation.
      Replacement of the two Ser residues at positions 32 and 36 corresponding to IKK
      phosphorylation sites by Ala resulted in almost complete prevention of binding of
      an IkappaBalpha-E3 to IkappaBalpha. These results indicate that phosphorylation
      of IkappaBalpha is necessary and sufficient for recruitment of this
      IkappaBalpha-E3 to associate with IkappaBalpha.
CI  - Copyright 1999 Academic Press.
FAU - Suzuki, H
AU  - Suzuki H
AD  - Institute for Drug Discovery Research, Yamanouchi Pharmaceutical Co., Ltd., 21
      Miyukigaoka, Ibaraki, Tsukuba-shi, 305-8585, Japan.
FAU - Chiba, T
AU  - Chiba T
FAU - Kobayashi, M
AU  - Kobayashi M
FAU - Takeuchi, M
AU  - Takeuchi M
FAU - Furuichi, K
AU  - Furuichi K
FAU - Tanaka, K
AU  - Tanaka K
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Biochem Biophys Res Commun
JT  - Biochemical and biophysical research communications
JID - 0372516
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (I-kappa B Proteins)
RN  - 0 (NF-kappa B)
RN  - 0 (NFKBIA protein, human)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Transcription Factor RelA)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 0 (Ubiquitins)
RN  - 139874-52-5 (NF-KappaB Inhibitor alpha)
RN  - 17885-08-4 (Phosphoserine)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 2.3.2.23 (Ubiquitin-Conjugating Enzymes)
RN  - EC 2.3.2.27 (Ubiquitin-Protein Ligases)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 2.7.11.10 (CHUK protein, human)
RN  - EC 2.7.11.10 (I-kappa B Kinase)
RN  - EC 2.7.11.10 (IKBKB protein, human)
RN  - EC 2.7.11.10 (IKBKE protein, human)
RN  - EC 6.- (Ligases)
SB  - IM
MH  - Adenosine Triphosphate/metabolism
MH  - Amino Acid Substitution
MH  - Animals
MH  - Blotting, Western
MH  - DNA-Binding Proteins/chemistry/genetics/*metabolism
MH  - Enzyme Activation/drug effects
MH  - HeLa Cells
MH  - Humans
MH  - I-kappa B Kinase
MH  - *I-kappa B Proteins
MH  - Ligases/*metabolism
MH  - NF-KappaB Inhibitor alpha
MH  - NF-kappa B/metabolism
MH  - Phosphorylation/drug effects
MH  - Phosphoserine/metabolism
MH  - Precipitin Tests
MH  - Protein Binding/drug effects
MH  - Protein-Serine-Threonine Kinases/*metabolism
MH  - Recombinant Fusion Proteins/chemistry/genetics/metabolism
MH  - Signal Transduction/drug effects
MH  - Time Factors
MH  - Transcription Factor RelA
MH  - Tumor Necrosis Factor-alpha/pharmacology
MH  - Ubiquitin-Conjugating Enzymes
MH  - Ubiquitin-Protein Ligases
MH  - Ubiquitins/*metabolism
EDAT- 1999/03/06 00:00
MHDA- 1999/03/06 00:01
CRDT- 1999/03/06 00:00
PHST- 1999/03/06 00:00 [pubmed]
PHST- 1999/03/06 00:01 [medline]
PHST- 1999/03/06 00:00 [entrez]
AID - S0006-291X(99)90296-6 [pii]
AID - 10.1006/bbrc.1999.0296 [doi]
PST - ppublish
SO  - Biochem Biophys Res Commun. 1999 Mar 5;256(1):121-6. doi: 10.1006/bbrc.1999.0296.