Domain structure of budding yeast CSN proteins and csn deletion phenotype(A) Schematic representation of budding yeast genes encoding proteins identified in a stable complex with Rri1p/Csn5p. Conserved MPN and PCI domains are highlighted as red and blue boxes, respectively. Consistent with the typical localization pointed out in the original report , the PCI domains of budding yeast CSN proteins are located in the C-terminus. The table to the right lists the ORF names next to published gene names, and names assigned by Glickman and colleagues (personal communication, denoted by *). (B) Summary of published budding yeast CSN protein interactions. Two-hybrid interactions [32,33] are indicated by dotted lines, in vivo protein interactions identified by affinity purification of Rri1p/Csn5p interacting proteins  are indicated by solid lines. (C) Effect of deletion of csn genes on Rub1p modification of Cdc53p. The indicated strains were grown in YPD media to an optical density of ~0.8 and total protein lysate was prepared by bead lysis in a buffer containing 20 mM Hepes pH 7.4, 150 mM NaCl, 0.2% Triton X-100, 1 mM EDTA, and 50 mM NaF, 1 mM DTT, 1 mM PMSF, 17 ug/ml aprotinin, 10 ug/ml leupeptin, and 10 ug/ml pepstatin. Lysates were separated on SDS gels and the native and Rub1p-modified forms of Cdc53p were detected by immunoblotting with Cdc53p antibodies (purchased from Santa Cruz Biotechnology, sc-6717). Unmodified and Rub1p-modified Cdc53p are indicated.
Wee et al. BMC Genetics 2002 3:15 doi:10.1186/1471-2156-3-15