DEGRADATION OF ORNITHINE DECARBOXYLASE BY THE MAMMALIAN AND YEAST 26S PROTEASOME COMPLEXES REQUIRES ALL THE COMPONENTS OF THE PROTEASE

被引:31
作者
ELIAS, S
BERCOVICH, B
KAHANA, C
COFFINO, P
FISCHER, M
HILT, W
WOLF, DH
CIECHANOVER, A
机构
[1] TECHNION ISRAEL INST TECHNOL, FAC MED, DEPT BIOCHEM, IL-31096 HAIFA, ISRAEL
[2] TECHNION ISRAEL INST TECHNOL, FAC MED, RAPPAPORT FAMILY INST RES MED SCI, IL-31096 HAIFA, ISRAEL
[3] WEIZMANN INST SCI, DEPT MOLEC GENET & VIROL, IL-76100 REHOVOT, ISRAEL
[4] UNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USA
[5] UNIV STUTTGART, INST BIOCHEM, W-7000 STUTTGART, GERMANY
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1995年 / 229卷 / 01期
关键词
2OS AND 26S PROTEASOME COMPLEXES; ORNITHINE DECARBOXYLASE; UBIQUITIN; DEGRADATION;
D O I
10.1111/j.1432-1033.1995.0276l.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Ornithine decarboxylase (ODC), a key enzyme in the biosynthesis of polyamines, is an extremely short-lived protein. This attribute is important for the regulation of the activity of the enzyme and implies that the mechanisms involved in its degradation play an important role in the control of the cellular processes in which the enzyme is involved. Recently, it has been shown that ODC is degraded by the 26S proteasome complex in a process that requires antizyme, but not ubiquitin. With one reported exception, ODC, the 26S complex recognizes and degrades specifically ubiquitinated proteins. Their unconjugated counterparts are not targeted. The 26S complex is composed of a core catalytic unit, the 20S proteasome complex, and two additional, and apparently distinct, subcomplexes. The two additional subcomplexes are regulatory subunits that are required in order to confer specificity and control. In this study, we demonstrate that, like the degradation of ubiquitin-conjugated proteins, ubiquitin-independent degradation of ODC also requires prior assembly of the mammalian 26S proteasome from all the three subunits, the 20S proteasome and the two subcomplexes. The combination of any two subunits does not support generation of a proteolytically active complex. This is also true for the yeast 26S complex. Like the mammalian 20S proteasome, the yeast 20S complex can cleave short peptides in an ATP-independent mode, but cannot degrade ODC or ubiquitin-conjugated proteins. These proteins are degraded only following addition of the regulatory subunits and generation of the high-molecular-mass 26S complex. In a distinct, but related, set of experiments, we demonstrate that the degradation of ODC by the assembled 26S proteasome in vitro reproduces faithfully proteolysis of the enzyme in the intact cell. Namely, (a) a C-terminal-deleted mouse ODC and trypanosome ODC are stable both in vitro and in vivo, and (b) like proteolysis in the intact cell, degradation in the reconstituted cell-free system is also dependent upon the addition of antizyme.
引用
收藏
页码:276 / 283
页数:8
相关论文
共 49 条
[1]  
BASS KE, 1992, J BIOL CHEM, V267, P11034
[2]   INVOLVEMENT OF THE 20S-PROTEASOME IN THE DEGRADATION OF ORNITHINE DECARBOXYLASE [J].
BERCOVICH, Z ;
KAHANA, C .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1993, 213 (01) :205-210
[3]  
BERCOVICH Z, 1989, J BIOL CHEM, V264, P15949
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[5]   DEGRADATION OF NUCLEAR ONCOPROTEINS BY THE UBIQUITIN SYSTEM INVITRO [J].
CIECHANOVER, A ;
DIGIUSEPPE, JA ;
BERCOVICH, B ;
ORIAN, A ;
RICHTER, JD ;
SCHWARTZ, AL ;
BRODEUR, GM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (01) :139-143
[6]   THE UBIQUITIN-PROTEASOME PROTEOLYTIC PATHWAY [J].
CIECHANOVER, A .
CELL, 1994, 79 (01) :13-21
[7]  
DEVERAUX Q, 1994, J BIOL CHEM, V269, P7059
[8]  
DUBIEL W, 1992, J BIOL CHEM, V267, P22699
[9]   ATP-DEPENDENT INCORPORATION OF 20S PROTEASE INTO THE 26S COMPLEX THAT DEGRADES PROTEINS CONJUGATED TO UBIQUITIN - (PROTEIN BREAKDOWN MULTICATALYTIC PROTEINASE COMPLEX) [J].
EYTAN, E ;
GANOTH, D ;
ARMON, T ;
HERSHKO, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (20) :7751-7755
[10]  
EYTAN E, 1993, J BIOL CHEM, V268, P4668