Rpn7 Is Required for the Structural Integrity of the 26 S Proteasome of Saccharomyces cerevisiae

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2004
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Saeki, Yasushi
Yokosawa, Hideyoshi
Toh-e, Akio
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The Journal of Biological Chemistry : JBC. 2004, 279(26), pp. 27168-27176. ISSN 0021-9258. eISSN 1083-351X. Available under: doi: 10.1074/jbc.M314231200
Zusammenfassung

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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570 Biowissenschaften, Biologie
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ISO 690ISONO, Erika, Yasushi SAEKI, Hideyoshi YOKOSAWA, Akio TOH-E, 2004. Rpn7 Is Required for the Structural Integrity of the 26 S Proteasome of Saccharomyces cerevisiae. In: The Journal of Biological Chemistry : JBC. 2004, 279(26), pp. 27168-27176. ISSN 0021-9258. eISSN 1083-351X. Available under: doi: 10.1074/jbc.M314231200
BibTex
@article{Isono2004-06-25Requi-42151,
  year={2004},
  doi={10.1074/jbc.M314231200},
  title={Rpn7 Is Required for the Structural Integrity of the 26 S Proteasome of Saccharomyces cerevisiae},
  number={26},
  volume={279},
  issn={0021-9258},
  journal={The Journal of Biological Chemistry : JBC},
  pages={27168--27176},
  author={Isono, Erika and Saeki, Yasushi and Yokosawa, Hideyoshi and Toh-e, Akio}
}
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