Additional file 2.

Figure S2. Analysis of Ddi1 and Dsk2 association with proteasomes isolated from rpn1-D517A , rpn10-uim rpn1-D571A, and rpn10-uim rpn13-KKD rpn1-D571A mutants. (A) Affinity-purified rpn1-D517A proteasomes contain reduced levels of Ddi1 and Ub conjugates. Levels of UBA-UBL proteins, the lid subunit Rpn12 and polyubiquitin are shown for affinity purified proteasomes (IP) and in the whole cell extract input (WCE). (B) Affinity-purified rpn10-uim rpn1-D517A proteasomes similarly show diminished association of Ddi1 and Ub conjugates compared to rpn10-uim proteasomes. (C) Affinity-purified rpn10-uim rpn13-KKD rpn1-D517A proteasomes contain reduced levels of Ddi1, Dsk2 and Ub conjugates in comparison to proteasomes from an rpn10-uim rpn13-KKD strain. Densitometric quantification of this blot is shown on the right. The amounts of UBL proteins were normalized to Rpn11FLAG and wild type levels were set as 100%.

Format: PDF Size: 728KB Download file

This file can be viewed with: Adobe Acrobat Reader

Gomez et al. BMC Biology 2011 9:33   doi:10.1186/1741-7007-9-33