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Functional Differences Of Two Forms Of The Inhibitor Of Caspase-activated DNase, ICAD-L, And ICAD-S*
H. Sakahira, M. Enari, S. Nagata
Published 1999 · Biology, Medicine
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Caspase-activated DNase (CAD) is responsible for the DNA fragmentation that occurs during apoptosis. CAD is complexed with an inhibitor of CAD (ICAD) in non-apoptotic, growing cells. Here, we report that mouse WR19L and human Jurkat T lymphoma cells express two alternative forms of ICAD, ICAD-L and ICAD-S, at similar levels. CAD was predominantly associated with ICAD-L in these cell lines. When CAD was expressed alone in Sf9 cells, it was found in insoluble fractions. However, when CAD was co-expressed with ICAD-L and ICAD-S, it was recovered as a soluble protein complexed predominantly with ICAD-L. In vitro transcription and translation of CAD cDNA did not produce a functional protein. Addition of ICAD-L but not ICAD-S to the assay mixture resulted in the synthesis of functional CAD. These results indicated that ICAD-L but not ICAD-S works as a specific chaperone for CAD, facilitating its correct folding during synthesis. Recombinant CAD, as a complex with ICAD-L, was then produced in Sf9 cells. The complex was treated with caspase 3, and CAD was purified to homogeneity. The purified CAD had DNase activity with a high specific activity.
This paper references
Calculation of protein extinction coefficients from amino acid sequence data.
S. Gill (1989)
Cell‐free synthesis and assembly of prolyl 4‐hydroxylase: the role of the beta‐subunit (PDI) in preventing misfolding and aggregation of the alpha‐subunit.
D. C. John (1993)
Substrate Specificities of Caspase Family Proteases*
R. Talanian (1997)
Cell death: the significance of apoptosis.
A. Wyllie (1980)
Small heat shock proteins are molecular chaperones.
U. Jakob (1993)
Nuclear changes in apoptosis.
W. Earnshaw (1995)
A matter of life and cell death.
G. Evan (1998)
Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation
A. Wyllie (1980)
The 40-kDa subunit of DNA fragmentation factor induces DNA fragmentation and chromatin condensation during apoptosis.
X. Liu (1998)
Reduced Chaperone-like Activity of αAins-crystallin, an Alternative Splicing Product Containing a Large Insert Peptide (*)
R. Smulders (1995)
Caspases: enemies within.
N. Thornberry (1998)
bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death
L. Boise (1993)
Developing Caenorhabditis elegans neurons may contain both cell-death protective and killer activities.
S. Shaham (1996)
Apoptosis: A Basic Biological Phenomenon with Wide-ranging Implications in Tissue Kinetics
J. Kerr (1972)
Apoptosis by Death Factor
S. Nagata (1997)
A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD
M. Enari (1998)
Involvement of MACH, a Novel MORT1/FADD-Interacting Protease, in Fas/APO-1- and TNF Receptor–Induced Cell Death
M. Boldin (1996)
Mitochondria and apoptosis.
D. Green (1998)
Ich-1, an Ice/ced-3-related gene, encodes both positive and negative regulators of programmed cell death
Lin Wang (1994)
Resistance to DNA fragmentation and chromatin condensation in mice lacking the DNA fragmentation factor 45.
J. Zhang (1998)
Site‐directed mutagenesis of human protein disulphide isomerase: effect on the assembly, activity and endoplasmic reticulum retention of human prolyl 4‐hydroxylase in Spodoptera frugiperda insect cells.
K. Vuori (1992)
Molecular cloning and characterization of human caspase-activated DNase.
Naomi Mukae (1998)
DFF, a Heterodimeric Protein That Functions Downstream of Caspase-3 to Trigger DNA Fragmentation during Apoptosis
X. Liu (1997)
Protein hydroxylation: prolyl 4‐hydroxylase, an enzyme with four cosubstrates and a multifunctional subunit
K. Kivirikko (1989)
Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis
H. Sakahira (1998)
Molecular chaperones in cellular protein folding
F. Hartl (1996)
Death receptors: signaling and modulation.
A. Ashkenazi (1998)
Cell suicide for beginners
M. Raff (1998)
Molecular chaperones in protein folding: the art of avoiding sticky situations.
F. Hartl (1994)
CPAN, a human nuclease regulated by the caspase-sensitive inhibitor DFF45
R. Halenbeck (1998)
Distinct signal transduction through the tyrosine‐containing domains of the granulocyte colony‐stimulating factor receptor.
A. Yoshikawa (1995)
A Combinatorial Approach Defines Specificities of Members of the Caspase Family and Granzyme B
N. Thornberry (1997)
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Identification and analysis of caspase substrates: proteolytic cleavage of poly(ADP-ribose)polymerase and DNA fragmentation factor 45.
C. Boucher (2001)
MiR-145, a new regulator of the DNA Fragmentation Factor-45 (DFF45)-mediated apoptotic network
J. Zhang (2010)
Purification, crystallization and preliminary X-ray crystallographic studies of Drep2 CIDE domain.
S. M. Lee (2014)
A new method for protein coexpression in Escherichia coli using two incompatible plasmids.
W. Yang (2001)
Inhibition of caspase-9 by oridonin, a diterpenoid isolated from Rabdosia rubescens, augments apoptosis in human laryngeal cancer cells
Ning Kang (2015)
General interaction mode of CIDE:CIDE complex revealed by a mutation study of the Drep2 CIDE domain
S. M. Lee (2013)
Elevated p53 expression is associated with dysregulation of the ubiquitin-proteasome system in dilated cardiomyopathy.
E. Birks (2008)
Nuclear transport of the DNA fragmentation factor via the classical importin α/β-pathway
Sonja Neimanis (2007)
A putative role of Drep1 in apoptotic DNA fragmentation system in fly is mediated by direct interaction with Drep2 and Drep4
O. K. Park (2013)
Caspase-3 Is the Primary Activator of Apoptotic DNA Fragmentation via DNA Fragmentation Factor-45/Inhibitor of Caspase-activated DNase Inactivation*
Beni B Wolf (1999)
Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling
Charles W Morgan (2015)
On the evolution of programmed cell death: apoptosis of the unicellular eukaryote Leishmania major involves cysteine proteinase activation and mitochondrion permeabilization
D. Arnoult (2002)
Activation of p53/p21/PUMA alliance and disruption of PI‐3/Akt in multimodal targeting of apoptotic signaling cascades in cervical cancer cells by a pentacyclic triterpenediol from Boswellia serrata
S. Bhushan (2009)
Crystallization and preliminary X-ray crystallographic studies of the CIDE-domain complex between Drep2 and Drep3 from Drosophila melanogaster.
Ok Kyeung Park (2011)
Physalin A induces apoptosis via p53-Noxa-mediated ROS generation, and autophagy plays a protective role against apoptosis through p38-NF-κB survival pathway in A375-S2 cells.
Hao He (2013)
A cyano analogue of boswellic acid induces crosstalk between p53/PUMA/Bax and telomerase that stages the human papillomavirus type 18 positive HeLa cells to apoptotic death.
Sheema Khan (2011)
Degradation of chromosomal DNA during apoptosis
S. Nagata (2003)
Specific Chaperone-like Activity of Inhibitor of Caspase-activated DNase for Caspase-activated DNase*
H. Sakahira (2000)
Interaction mode of CIDE family proteins in fly: DREP1 and DREP3 acidic surfaces interact with DREP2 and DREP4 basic surfaces
C. M. Kim (2017)
Loss of splicing factor ASF/SF2 induces G2 cell cycle arrest and apoptosis, but inhibits internucleosomal DNA fragmentation.
X. Li (2005)
Regulation of DNA fragmentation: the role of caspases and phosphorylation
Ikuko Kitazumi (2011)
Tetrahydrocurcumin induces G2/M cell cycle arrest and apoptosis involving p38 MAPK activation in human breast cancer cells.
N. Kang (2014)
Mechanism of nitric oxide‐induced apoptosis in human neuroblastoma SH‐SY5Y cells
R. Moriya (2000)
Enzymatic active site of caspase-activated DNase (CAD) and its inhibition by inhibitor of CAD.
H. Sakahira (2001)
Cleavage Preferences of the Apoptotic Endonuclease DFF40 (Caspase-activated DNase or Nuclease) on Naked DNA and Chromatin Substrates*
P. Widłak (2000)
Breakdown of Chromosomal DNA
S. Nagata (2002)
Activation of extracellular signal‐regulated kinases ERK1 and ERK2 induces Bcl‐xL up‐regulation via inhibition of caspase activities in erythropoietin signaling
M. Mori (2003)
Mathematical Modeling of Caspase Activated DNAase (CAD) Regulation Towards Apoptosis
M. Baharuddin (2008)
A triterpenediol from Boswellia serrata induces apoptosis through both the intrinsic and extrinsic apoptotic pathways in human leukemia HL-60 cells
S. Bhushan (2007)
Alternative Splicing and Control of Apoptotic DNA Fragmentation
X. Li (2006)
Structural mechanism for inactivation and activation of CAD/DFF40 in the apoptotic pathway.
Eui-Jeon Woo (2004)
A Novel Activation Mechanism of Caspase-activated DNase fromDrosophila melanogaster*
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