Nucling recruits Apaf-1/pro-caspase-9 complex for the induction of stress-induced apoptosis

Takashi Sakai; Li Liu; Xichuan Teng; Rika Mukai-Sakai; Hidenori Shimada; Ryuji Kaji; Tasuku Mitani; Mitsuru Matsumoto; Kazunori Toida; Kazunori Ishimura; Yuji Shishido; Tak W Mak; Kiyoshi Fukui

doi:10.1074/jbc.M402902200

Nucling recruits Apaf-1/pro-caspase-9 complex for the induction of stress-induced apoptosis

J Biol Chem. 2004 Sep 24;279(39):41131-40. doi: 10.1074/jbc.M402902200. Epub 2004 Jul 22.

Authors

Takashi Sakai¹, Li Liu, Xichuan Teng, Rika Mukai-Sakai, Hidenori Shimada, Ryuji Kaji, Tasuku Mitani, Mitsuru Matsumoto, Kazunori Toida, Kazunori Ishimura, Yuji Shishido, Tak W Mak, Kiyoshi Fukui

Affiliation

¹ The Institute for Enzyme Research, The University of Tokushima, Tokushima 770-8503, Japan.

PMID: 15271982
DOI: 10.1074/jbc.M402902200

Abstract

Nucling is a novel protein isolated from murine embryonal carcinoma cells with an up-regulated expression during cardiac muscle differentiation. We show here that Nucling was up-regulated by proapoptotic stimuli and important for the induction of apoptosis after cytotoxic stress. We further demonstrated that overexpressed Nucling was able to induce apoptosis. In Nucling-deficient cells, the expression levels of Apaf-1 and cytochrome c, which are the major components of an apoptosis-promoting complex named apoptosome, were both down-regulated under cellular stress. A deficiency of Nucling also conferred resistance to apoptotic stress on the cell. After UV irradiation, Nucling was shown to reside in an Apaf-1/pro-caspase-9 complex, suggesting that Nucling might be a key molecule for the formation and maintenance of this complex. Nucling induced translocation of Apaf-1 to the nucleus, thereby distributing the Nucling/Apaf-1/pro-caspase-9 complex to the nuclear fraction. These findings suggest that Nucling recruits and transports the apoptosome complex during stress-induced apoptosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alleles
Amino Acid Chloromethyl Ketones / chemistry
Animals
Apoptosis*
Apoptotic Protease-Activating Factor 1
Blotting, Northern
Blotting, Western
COS Cells
Caspase 9
Caspases / metabolism
Cell Death
Cell Line
Cysteine Proteinase Inhibitors / pharmacology
Cytochromes c / metabolism
Dose-Response Relationship, Drug
Down-Regulation
Electrophoresis, Gel, Two-Dimensional
Electrophoresis, Polyacrylamide Gel
Genetic Vectors
HeLa Cells
Humans
Hydrogen Peroxide / chemistry
In Situ Nick-End Labeling
Membrane Proteins / physiology*
Mice
Mice, Transgenic
Microscopy, Confocal
Mitochondria / metabolism
Models, Genetic
Plasmids / metabolism
Proteins / metabolism*
RNA / chemistry
Reverse Transcriptase Polymerase Chain Reaction
Transfection
Transgenes
Ultraviolet Rays
Up-Regulation

Substances

APAF1 protein, human
Amino Acid Chloromethyl Ketones
Apaf1 protein, mouse
Apoptotic Protease-Activating Factor 1
Cysteine Proteinase Inhibitors
Membrane Proteins
Proteins
Uaca protein, mouse
benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
RNA
Cytochromes c
Hydrogen Peroxide
CASP9 protein, human
Casp9 protein, mouse
Caspase 9
Caspases