Article
- The EMBO Journal (2004) 23, 500 - 510
- doi:10.1038/sj.emboj.7600059
Published online: 22 January 2004
Subject Categories:
Plasticity in eucaryotic 20S proteasome ring assembly revealed by a subunit deletion in yeast
Irina Velichutinaa, Pamela L Connerlyab, Cassandra S Arendtc, Xia Li and Mark Hochstrasser
- Department of Molecular Biophysics & Biochemistry, Yale University, New Haven, CT, USA
Correspondence to:
Mark Hochstrasser, Department of Molecular Biophysics & Biochemistry, Yale University, 266 Whitney Avenue, PO Box 208114, New Haven, CT 06520, USA. Tel.: +1 203 432 5101; Fax: +1 203 432 5175; E-mail: mark.hochstrasser@yale.edu
aThese authors contributed equally to the work
bCurrent address: Department of Microbiology, Miami University, 32 Pearson Hall, Oxford, OH 45056, USA
cCurrent address: Department of Biochemistry & Molecular Biology, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA
Received 21 October 2003; Accepted 8 December 2003
Abstract
The 20S proteasome is made up of four stacked heptameric rings, which in eucaryotes assemble from 14 different but related subunits. The rules governing subunit assembly and placement are not understood. We show that a different kind of proteasome forms in yeast when the Pre9/
3 subunit is deleted. Purified pre9
proteasomes show a two-fold enrichment for the Pre6/4 subunit, consistent with the presence of an extra copy of Pre6 in each outer ring. Based on disulfide engineering and structure-guided suppressor analyses, Pre6 takes the position normally occupied by Pre9, a substitution that depends on a network of intersubunit salt bridges. When Arabidopsis PAD1/4 is expressed in yeast, it complements not only pre6 but also pre6 pre9 mutants; therefore, the plant 4 subunit also can occupy multiple positions in a functional yeast proteasome. Importantly, biogenesis of proteasomes is delayed at an early stage in pre9 cells, suggesting an advantage for Pre9 over Pre6 incorporation at the 3 position that facilitates correct assembly.
Keywords:
- proteasome,
- protein assembly,
- proteolysis,
- ubiquitin
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