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Molecular and Cellular Biology, October 1999, p. 7216-7227, Vol. 19, No. 10
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

ATP-Dependent Inactivation and Sequestration of Ornithine Decarboxylase by the 26S Proteasome Are Prerequisites for Degradation

Yasuko Murakami,1,* Senya Matsufuji,1 Shin-Ichi Hayashi,1 Nobuyuki Tanahashi,2 and Keiji Tanaka2

Department of Biochemistry 2, Jikei University School of Medicine, Minato-ku, Tokyo 105-8461,1 and The Tokyo Metropolitan Institute of Medical Science and CREST, Japan Science and Technology Corporation, Bunkyo-ku, Tokyo 113-8613,2 Japan

Received 6 January 1999/Returned for modification 24 February 1999/Accepted 19 July 1999

The 26S proteasome is a eukaryotic ATP-dependent protease, but the molecular basis of its energy requirement is largely unknown. Ornithine decarboxylase (ODC) is the only known enzyme to be degraded by the 26S proteasome without ubiquitinylation. We report here that the 26S proteasome is responsible for the irreversible inactivation coupled to sequestration of ODC, a process requiring ATP and antizyme (AZ) but not proteolytic activity. Neither the 20S proteasome (catalytic core) nor PA700 (the regulatory complex) by itself contributed to this ODC inactivation. Analysis with a C-terminal mutant ODC revealed that the 26S proteasome recognizes the C-terminal degradation signal of ODC exposed by attachment of AZ, and subsequent ATP-dependent sequestration of ODC in the 26S proteasome causes irreversible inactivation, possibly unfolding, of ODC and dissociation of AZ. These processes may be linked to the translocation of ODC into the 20S proteasomal inner cavity, centralized within the 26S proteasome, for degradation.

* Corresponding author. Mailing address: Department of Biochemistry 2, Jikei University School of Medicine, 3-25-8 Nishi-Shinbashi, Minato-ku, Tokyo 105-8461, Japan. Phone: 81-3-3433-1111. Fax: 81-3-3436-3897. E-mail: yasukomu{at}jikei.ac.jp.

Molecular and Cellular Biology, October 1999, p. 7216-7227, Vol. 19, No. 10
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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