ced-9, a member of the bcl-2 gene family in Caenorhabditis elegans plays a central roles in preventing cell death in worms. Overexpression of human bcl-2 can partially prevent cell death in C. elegans. However, it remains to be elucidated whether ced-9 can regulate cell death when expressed in other organisms. We demonstrated that the CED-9 protein is co-localized with BCL-2 in COS cells and Drosophila Schneider's L2 (SL2) cells, suggesting that the site of CED-9 action is located to specific cytoplasmic compartments. Overexpression of ced-9 only poorly protected cells from the death induced by ced-3 in HeLa cells, but ced-9 significantly reduced the cell death induced by ced-3 in Drosophila SL2 cells. Furthermore, apoptosis of SL2 cells that was induced by a Drosophila cell-death gene, reaper, was shown to be partially prevented by ced-9, bcl-2 and bcl-xL. These results suggest that the signaling pathway that is required for the anti-apoptotic function of bcl-2 family members, including ced-9, is conserved in Drosophila cells. In addition, SL2 cells provide a unique systems for dissecting the main machinery of cell death.
REFERENCES
Beidler
D. R.
, Tewari
M.
, Friesen
P. D.
, Poirier
G.
, Dixit
V. M.
(1995
). The baculovirus p35 protein inhibits Fas-and tumor necrosis factor-induced apoptosis.
J. Biol. Chem
270
, 16526
–16528
Ferenz
C.
, Franklin
S.
, Ghayur
T.
, Li
P.
, Licari
P.
, Mankovich
J.
, Shi
L.
, Greenberg
A. H.
, Miller
L. K.
, Wong
W. W.
(1995
). Inhibition of ICE family proteases by baculovirus antiapoptotic protein p35.
Science
269
, 1885
–1888
Chen
P.
, Nordstrom
W.
, Gish
B.
, Abrams
J. M.
(1996
). grim, a novel cell death gene in Drosophila.
Genes Dev
10
, 1773
–1782
Chinnaiyan
A. M.
, O'Rourke
K.
, Lane
B. R.
, Dixit
V. M.
(1997
). Interaction of CED-4 with CED-3 and CED-9: a molecular framework for cell death.
Science
275
, 1122
–1126
Chinnaiyan
A. M.
, Chaudhary
D.
, O'Rourke
K.
, Koonin
E. V.
, Dixit
V. M.
(1997
). Role of CED-4 in the activation of CED-3.
Nature
388
, 728
–729
Dolle
R. E.
, Hoyer
D.
, Prasad
C. V. C.
, Schmidt
S. J.
, Helaszek
C. T.
, Miller
R. E.
, Ator
M. A.
(1994
). Aspartate-based peptide a-((2,6-dichlorobenzoyl)oxy)methyl ketones as potent time-dependent inhibitors of interleukin-1-converting enzyme.
J. Med. Chem
37
, 563
–564
Durfee
T.
, Becherer
K.
, Chen
P. L.
, Yeh
S. H.
, Yang
Y.
, Kilbburn
A. E.
, Lee
W. H.
, Elledge
S. J.
(1993
). The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit.
Genes Dev
7
, 555
–569
Ellis
H. M.
, Horvitz
H. R.
(1986
). Genetic control of programmed cell death in the nematode C. elegans.
Cell
44
, 817
–829
Fraser
A. G.
, Evan
G. I.
(1997
). Identification of a Drosophila melanogaster ICE/CED-3-related protease, drICE.
EMBO J
16
, 2805
–2813
Gagliardini
V.
, Fernandez
P.-A.
, Lee
R. K. K.
, Drexler
H. C. A.
, Rotello
R. J.
, Fishman
M. C.
, Yuan
J.
(1994
). Prevention of vertebrate neuronal death by the crmA gene.
Science
263
, 826
–828
Grether
M. E.
, Abrams
J. M.
, Agapite
J.
, White
K.
, Steller
H.
(1995
). The head involution defective gene of Drosophila melanogaster functions in programmed cell death.
Genes Dev
9
, 1694
–1708
Hay
B. A.
, Wassarman
D. A.
, Rubin
G. M.
(1995
). Drosophila homologs of baculovirus inhibitor of apoptosis proteins function to block cell death.
Cell
83
, 1253
–1262
Hengartner
M. O.
, Horvitz
H. R.
(1994
). Activation of C. elegans cell death protein CED-9 by an amino-acid substitution in a domain converted in Bcl-2.
Nature
369
, 318
–320
Hisahara
S.
, Shoji
S.
, Okano
H.
, Miura
M.
(1997
). ICE/CED-3 family executes oligodendrocyte apoptosis by tumor necrosis factor.
J. Neurochem
69
, 10
–20
Jacobson
M. D.
, Burne
J. F.
, King
M. P.
, Miyashita
T.
, Reed
J. C.
, Raff
M. C.
(1993
). Bcl-2 blocks apoptosis in cells lacking mitochondrial DNA.
Nature
361
, 365
–368
Kluck
R. M.
, Bossy-Wetzel
E.
, Green
D. R.
, Newmeyer
D. D.
(1997
). The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis.
Science
275
, 1132
–1136
Komiyama
T.
, Ray
C. A.
, Pickup
D. J.
, Howard
A. D.
, Thornberry
N. A.
, Petersen
E. P.
, Salvesen
G.
(1994
). Inhibition of interleukin-1converting enzyme by the cowpox virus serpin CrmA. An example of cross-class inhibition.
J. Biol. Chem
269
, 19331
–19337
Krajewski
S.
, Tanaka
S.
, Takayama
S.
, Schneider
M. J.
, Fenton
W.
, Reed
J. C.
(1993
). Investigations of the subcellular distribution of the BCL-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membranes.
Cancer Res
53
, 4701
–4714
Minn
A. J.
, Velez
P.
, Schendel
S. L.
, Liang
H.
, Muchmore
S. W.
, Fesik
S. W.
, Fill
M.
, Thompson
C. B.
(1997
). Bcl-xL forms an ion channel in synthetic lipid membranes.
Nature
385
, 353
–357
Miura
M.
, Zhu
H.
, Rotello
R.
, Hartwieg
E. A.
, Yuan
J.
(1993
). Induction of apoptosis in fibroblastts by IL-1-converting enzyme, a mammalian homolog of the C. elegans cell death gene ced-3.
Cell
75
, 653
–660
Miura
M.
, Yuan
J.
(1996
). Mechanism of programmed cell death in Caenorhabditis elegans and vertebrates.
Curr. Top. Dev. Biol
32
, 139
–174
Nguyen
M.
, Branton
P.
, Walton
P.
, Oltvai
P.
, Korsmeyer
S.
, Shore
G.
(1994
). Role of membrane anchor domain of bcl-2 in suppression of apoptosis caused by E1B-defective adenovirus.
J. Biol. Chem
269
, 16521
–16524
Pronk
G. J.
, Ramer
K.
, Amiri
P.
, Williams
L. T.
(1996
). Requirement of an ICE-like protease for induction of apoptosis and ceramide generation by REAPER.
Science
271
, 808
–810
Schendel
S. L.
, Xie
Z.
, Montal
M. O.
, Matsuyama
S.
, Reed
J. C.
(1997
). Channel formation by antiapoptotic protein bcl-2.
Proc. Nat. Acad. Sci. USA
94
, 5113
–5118
Schneider
I.
(1972
). Cell lines derived from the late embryonic stages of Drosophila melanogaster.
J. Embryol. Exp. Morphol
27
, 353
–356
Seshagiri
S.
, Miller
L. K.
(1997
). Caenorhabditis elegans CED-4 stimulates CED-3 processing and CED-3-induced apoptosis.
Curr. Biol
7
, 455
–460
Shaham
S.
, Horvitz
H. R.
(1996
). An alternatively spliced C. elegans ced-4 RNA encodes a novel cell death inhibitor.
Cell
86
, 201
–208
Shaham
S.
, Horvitz
R.
(1996
). Developing Caenorhabditis elegans neurons may contain both cell-death protective and killer activities.
Genes Dev
10
, 578
–591
Shibasaki
F.
, Kondo
E.
, Akagi
T.
, Mckeon
F.
(1997
). Suppression of signalling through NF-AT by interactions between calcineurin and bcl-2.
Nature
386
, 728
–731
Song
Z.
, McCall
K.
, Steller
H.
(1997
). DCP-1, a Drosophila cell death protease essential for development.
Science
275
, 536
–540
Sugimoto
A.
, Friesen
P. D.
, Rothman
J. H.
(1994
). Baculovirus p35 prevents developmentally programmed cell death and rescues a ced-9 mutant in the nematode Caenorhabditis elegans.
EMBO J
13
, 2023
–2028
Susin
S. A.
, Zamzami
N.
, Castedo
M.
, Hirsch
T.
, Marchetti
P.
, Macho
A.
, Daugas
E.
, Geuskens
M.
, Kroemer
G.
(1996
). Bcl-2 inhibits the mitochondrial release of an apoptogenic protease.
J. Exp. Med
184
, 1331
–1341
Takahashi
A.
, Earnshaw
W. C.
(1996
). ICE-related proteases in apoptosis.
Curr. Opin. Genet. Dev
6
, 50
–55
Takayama
S.
, Sato
T.
, Krajewski
S.
, Kochel
K.
, Irie
S.
, Millan
J. A.
, Reed
J. C.
(1995
). Cloning and functional analysis of BAG-1: a novel Bcl-2 binding protein with anti-cell death activity.
Cell
80
, 279
–284
Tanaka
S.
, Saito
K.
, Reed
J. C.
(1993
). Structure-function analysis of the apoptosis-supressing Bcl-2 oncoprotein: substitution of a heterologous transmembrane domain restores function to truncated Bcl-2 protein.
J. Biol. Chem
268
, 10920
–10926
Wang
H.-G.
, Rapp
U. R.
, Reed
J. C.
(1996
). Bcl-2 targets the protein kinase Raf-1 to mitochondria.
Cell
87
, 629
–638
White
K.
, Grether
M. E.
, Abrams
J. M.
, Young
L.
, Farrell
K.
, Steller
H.
(1994
). Genetic control of programmed cell death in Drosophila.
Science
264
, 677
–683
White
K.
, Tahaoglu
E.
, Steller
H.
(1996
). Cell killing by the Drosophila gene reaper.
Science
271
, 805
–807
Wu
D.
, Wallen
H. D.
, Nunez
G.
(1997
). Interaction and regulation of subcellular localization of CED-4 by CED-9.
Science
275
, 1126
–1129
Wu
D.
, Wallen
H. D.
, Inohara
N.
, Nunez
G.
(1997
). Interaction and regulation of the Caenorhabditis elegans death protease CED-3 by CED-4 and CED-9.
J. Biol. Chem
272
, 21449
–21454
Xue
D.
, Horvitz
R.
(1995
). Inhibition of the Caenorhabditis elegans cell-death protease CED-3 by a CED-3 cleavage site in baculovirus p35 protein.
Nature
377
, 248
–251
Yang
E.
, Korsmeyer
S. J.
(1996
). Molecular thanatopsis: a discourse on the BCL2 family and cell death.
Blood
88
, 386
–401
Yang
J.
, Liu
X.
, Bhalla
K.
, Kim
C. N.
, Ibrado
A. M.
, Cai
J.
, Peng
T.-I.
, Jones
D. P.
, Wang
X.
(1997
). Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked.
Science
275
, 1129
–1132
Yin
X. M.
, Oltvai
Z. N.
, Korsmeyer
S. J.
(1994
). BH1 and BH2 domains of Bcl-2 are required for inhibition of apoptosis and heterodimerization with Bax.
Nature
369
, 321
–323
Yuan
J.
, Horvitz
H. R.
(1992
). The Caenorhabditis elegans cell death gene ced-4 encodes a novel protein and is expressed during the period of extensive programmed cell death.
Development
116
, 309
–320
Yuan
J.
, Shaham
S.
, Ledoux
S.
, Ellis
H. M.
, Horovitz
H. R.
(1993
). The C. elegans cell death gene ced-3 encodes a protein similar to mammalian interleukin-1-converting enzyme.
Cell
75
, 641
–652
Zha
H.
, Aime-Sempe
C.
, Sato
T.
, Reed
J. C.
(1996
). Proapoptotic protein Bax heterodimerizes with Bcl-2 amd homodimerizes with Bax via a novel domain (BH3) distinct from BH1 and BH2.
J. Biol. Chem
271
, 7440
–7444
Zou
H.
, Henzel
W. J.
, Liu
W.
, Lutschg
A.
, Wang
X.
(1997
). Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3.
Cell
90
, 405
–413
This content is only available via PDF.
© 1998 by Company of Biologists
1998
