Rpn7 is required for the structural integrity of the 26 S proteasome of Saccharomyces cerevisiae

被引:40
作者
Isono, E
Saeki, Y
Yokosawa, H
Toh-e, A [1 ]
机构
[1] Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Tokyo 1130033, Japan
[2] Hokkaido Univ, Grad Sch Pharmaceut Sci, Dept Biochem, Sapporo, Hokkaido 0600812, Japan
关键词
D O I
10.1074/jbc.M314231200
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
Rpn7 is one of the lid subunits of the 26 S proteasome regulatory particle. The RPN7 gene is known to be essential, but its function remains to be elucidated. To explore the function of Rpn7, we isolated and characterized temperature-sensitive rpn7 mutants. All of the rpn7 mutants obtained accumulated poly-ubiquitinated proteins when grown at the restrictive temperature. The N-end rule substrate (Ub-Arg-beta-galactosidase), the UFD pathway substrate ( Ub-Pro-beta-galactosidase), and cell cycle regulators (Pds1 and Clb2) were found to be stabilized in experiments using one of the rpn7 mutants termed rpn7-3 at the restrictive temperature, indicating its defect in the ubiquitin-proteasome pathway. Subsequent analysis of the structure of the 26 S proteasome in rpn7-3 cells suggested that the defect was in the assembly of the 26 S holoenzyme. The most striking characteristic of the proteasome of the rpn7-3 mutant was that a lid subcomplex affinity-purified from the rpn7-3 cells grown at the restrictive temperature contained only 5 of the 8 lid components, a phenomenon that has not been reported in the previously isolated lid mutants. From these results, we concluded that Rpn7 is required for the integrity of the 26 S complex by establishing a correct lid structure.
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页码:27168 / 27176
页数:9
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