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J. Biol. Chem., Vol. 277, Issue 5, 3364-3370, February 1, 2002
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From the Department of Genetics, Osaka University Medical School,
and Core Research for Evolutional Science and Technology, Japan Science
and Technology Corporation, 2-2 Yamada-oka, Suita, Osaka
565-0871, Japan
CAD (caspase-activated DNase) that causes
chromosomal DNA fragmentation during apoptosis exists as a complex with
ICAD (inhibitor of CAD) in proliferating cells. Here, we report that
denatured CAD is functionally refolded with Hsc70-Hsp40 and ICAD.
Hsc70-Hsp40 suppresses the aggregation of the denatured CAD, but cannot
restore its enzymatic activity. In contrast, ICAD could not suppress
the aggregation of CAD, but supported the CAD's renaturation with Hsc70-Hsp40, indicating that ICAD recognizes the
quasi-native folding state of CAD that is conferred by
Hsc70-Hsp40. Using an in vitro translation system, we then
showed that during CAD translation, Hsc70-Hsp40 as well as ICAD bind to
the nascent CAD polypeptide, while on ribosomes. These results indicate
that ICAD together with Hsc70-Hsp40 assists the folding of CAD during
its synthesis, and that the CAD·ICAD heterodimer is formed
co-translationally.
Co-translational Folding of Caspase-activated DNase with Hsp70,
Hsp40, and Inhibitor of Caspase-activated DNase*
and
*
This work was supported in part by grants-in-aid from the
Ministry of Education, Science, Sports, and Culture in Japan.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Supported by Research Fellowships of the Japan Society for the
Promotion of Science. Present address: Max-Planck-Institute for
Biochemistry, Dept. of Cellular Biochemistry, Am Klopferspitz 18a,
D-82152 Martinsried, Germany.
§
To whom coorespondence should be addressed: Dept. of Genetics,
Osaka University Medical School, B-3, 2-2 Yamada-oka, Suita, Osaka
565-0871, Japan. Tel.: 81-6-6879-3310; Fax: 81-6-6879-3319; E-mail:
nagata@genetic.med.osaka-u.ac.jp.
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