PMID- 10393175
OWN - NLM
STAT- MEDLINE
DCOM- 19990831
LR  - 20161124
IS  - 0261-4189 (Print)
IS  - 0261-4189 (Linking)
VI  - 18
IP  - 13
DP  - 1999 Jul 1
TI  - Role of cytochrome c and dATP/ATP hydrolysis in Apaf-1-mediated caspase-9
      activation and apoptosis.
PG  - 3586-95
AB  - Apaf-1 plays a critical role in apoptosis by binding to and activating
      procaspase-9. We have identified a novel Apaf-1 cDNA encoding a protein of 1248
      amino acids containing an insertion of 11 residues between the CARD and ATPase
      domains, and another 43 amino acid insertion creating an additional WD-40 repeat.
      The product of this Apaf-1 cDNA activated procaspase-9 in a cytochrome c and
      dATP/ATP-dependent manner. We used this Apaf-1 to show that Apaf-1 requires
      dATP/ATP hydrolysis to interact with cytochrome c, self-associate and bind to
      procaspase-9. A P-loop mutant (Apaf-1K160R) was unable to associate with Apaf-1
      or bind to procaspase-9. Mutation of Met368 to Leu enabled Apaf-1 to
      self-associate and bind procaspase-9 independent of cytochrome c, though still
      requiring dATP/ATP for these activities. The Apaf-1M368L mutant exhibited greater
      ability to induce apoptosis compared with the wild-type Apaf-1. We also show that
      procaspase-9 can recruit procaspase-3 to the Apaf-1-procaspase-9 complex.
      Apaf-1(1-570), a mutant lacking the WD-40 repeats, associated with and activated 
      procaspase-9, but failed to recruit procaspase-3 and induce apoptosis. These
      results suggest that the WD-40 repeats may be involved in procaspase-9-mediated
      procaspase-3 recruitment. These studies elucidate biochemical steps required for 
      Apaf-1 to activate procaspase-9 and induce apoptosis.
FAU - Hu, Y
AU  - Hu Y
AD  - Department of Pathology and Comprehensive Cancer Center, University of Michigan
      Medical School, 1500 East Medical Center Drive, 4219 CCGC, Ann Arbor, MI 48109,
      USA.
FAU - Benedict, M A
AU  - Benedict MA
FAU - Ding, L
AU  - Ding L
FAU - Nunez, G
AU  - Nunez G
LA  - eng
GR  - 2T32HL07517/HL/NHLBI NIH HHS/United States
GR  - CA-64421/CA/NCI NIH HHS/United States
GR  - CA-64556/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - EMBO J
JT  - The EMBO journal
JID - 8208664
RN  - 0 (APAF1 protein, human)
RN  - 0 (Apoptotic Protease-Activating Factor 1)
RN  - 0 (Cytochrome c Group)
RN  - 0 (Deoxyadenine Nucleotides)
RN  - 0 (Enzyme Precursors)
RN  - 0 (Proteins)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 3.4.22.- (CASP3 protein, human)
RN  - EC 3.4.22.- (CASP9 protein, human)
RN  - EC 3.4.22.- (Caspase 3)
RN  - EC 3.4.22.- (Caspase 9)
RN  - EC 3.4.22.- (Caspases)
RN  - K8KCC8SH6N (2'-deoxyadenosine triphosphate)
SB  - IM
MH  - Adenosine Triphosphate/*metabolism
MH  - Alternative Splicing
MH  - Amino Acid Substitution
MH  - *Apoptosis
MH  - Apoptotic Protease-Activating Factor 1
MH  - Caspase 3
MH  - Caspase 9
MH  - Caspases/*metabolism
MH  - Cell Line
MH  - Cytochrome c Group/*metabolism
MH  - Deoxyadenine Nucleotides/*metabolism
MH  - Enzyme Activation
MH  - Enzyme Precursors/*metabolism
MH  - Humans
MH  - Hydrolysis
MH  - Models, Biological
MH  - Mutation
MH  - Protein Binding
MH  - Proteins/chemistry/genetics/*metabolism
MH  - Time Factors
PMC - PMC1171437
EDAT- 1999/07/07 00:00
MHDA- 1999/07/07 00:01
CRDT- 1999/07/07 00:00
PHST- 1999/07/07 00:00 [pubmed]
PHST- 1999/07/07 00:01 [medline]
PHST- 1999/07/07 00:00 [entrez]
AID - 10.1093/emboj/18.13.3586 [doi]
PST - ppublish
SO  - EMBO J. 1999 Jul 1;18(13):3586-95. doi: 10.1093/emboj/18.13.3586.