TY - JOUR
T1 - Actin cleavage by CPP-32/apopain during the development of apoptosis
AU - Mashima, Tetsuo
AU - Naito, Mikihiko
AU - Noguchi, Kohji
AU - Miller, Douglas K.
AU - Nicholson, Donald W.
AU - Tsuruo, Takashi
N1 - Funding Information:
We thank Drs H Kawai and S Kataoka for providing us with the inhibitors of ICE family proteases. We also thank Dr T Akizawa for synthesis of GQVITC peptide under the support of a special grant for Advanced Research on Cancer. We are grateful to Drs A Tomida, N Fujita, H Seimiya and N Haga for helpful discussion. This work was supported by a special grant for Advanced Research on Cancer, and Grants-in-Aid for Cancer Research from the Ministry of Education, Science and Culture, Japan. TM received the Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists.
PY - 1997
Y1 - 1997
N2 - Interleukin-1β-converting enzyme (ICE)/ced-3 family proteases play key roles in apoptosis. However, cellular substrates for ICE family proteases involved in apoptosis are not well understood. We previously showed that actin is cleaved ill vitro by an ICE family protease, distinct from ICE itself, which is activated during VP-16-induced apoptosis. In this report, we demonstrate that the actin-cleaving ICE-family protease in the apoptotic cell extract is the activated CPP-32/apopain. CPP-32 effectively cleaves actin protein to 15 kDa and 31 kDa fragments. Studies with an antibody raised against Gly-Gln-Val-IIe-Thr peptide, the N-terminal sequence of the cleaved 15 kDa actin fragment, showed that actin is also cleaved in vivo during the development of apoptosis. Moreover, Benzyloxycarbonyl-Glu-Val-Asp-CH2OC(O)-2,6,-dichlorobenzene (Z-EVD-CH2-DCB), a selective inhibitor of CPP-32(-Like) protease, efficiently inhibited the cleavage of actin and the apoptosis of VP-16-treated U937 cells. Our present results indicate that actin is the substrate of CPP-32/apopain(-like) protease both in vitro and in vivo and suggest the role of actin in the control of cell growth and apoptosis.
AB - Interleukin-1β-converting enzyme (ICE)/ced-3 family proteases play key roles in apoptosis. However, cellular substrates for ICE family proteases involved in apoptosis are not well understood. We previously showed that actin is cleaved ill vitro by an ICE family protease, distinct from ICE itself, which is activated during VP-16-induced apoptosis. In this report, we demonstrate that the actin-cleaving ICE-family protease in the apoptotic cell extract is the activated CPP-32/apopain. CPP-32 effectively cleaves actin protein to 15 kDa and 31 kDa fragments. Studies with an antibody raised against Gly-Gln-Val-IIe-Thr peptide, the N-terminal sequence of the cleaved 15 kDa actin fragment, showed that actin is also cleaved in vivo during the development of apoptosis. Moreover, Benzyloxycarbonyl-Glu-Val-Asp-CH2OC(O)-2,6,-dichlorobenzene (Z-EVD-CH2-DCB), a selective inhibitor of CPP-32(-Like) protease, efficiently inhibited the cleavage of actin and the apoptosis of VP-16-treated U937 cells. Our present results indicate that actin is the substrate of CPP-32/apopain(-like) protease both in vitro and in vivo and suggest the role of actin in the control of cell growth and apoptosis.
KW - Actin
KW - Apoptosis
KW - CPP-32/apopain
KW - ICE
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U2 - 10.1038/sj.onc.1200919
DO - 10.1038/sj.onc.1200919
M3 - Article
C2 - 9070648
AN - SCOPUS:0030888898
SN - 0950-9232
VL - 14
SP - 1007
EP - 1012
JO - Oncogene
JF - Oncogene
IS - 9
ER -
