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. Author manuscript; available in PMC: 2015 Jul 17.
Published in final edited form as: Cell. 2014 Jul 17;158(2):434–448. doi: 10.1016/j.cell.2014.05.039

Figure 7. NUDC family proteins are specific co-chaperones for β-propeller domains.

Figure 7

A) NUDC family co-chaperones, SGT1, and CDC37 were assayed with a quantitative LUMIER assay for interaction with 80 kinases, 156 LRR domain proteins, and 275 proteins with a β-propeller domain. Bait proteins are organized by domain (annotated below) and rank-sorted based on their interaction with a specific co-chaperone (kinases with CDC37, LRRs with SGT1, WD40 with NUDC, RCC1 with NUDCD2, Kelch with NUDCD3).

B) Co-chaperones recognize specific domains in their clients. Indicated full-length proteins or truncated constructs were tagged with a 3xFLAG epitope and transfected into 293T cells. Their interaction with endogenous Hsp90 (top panel) or with endogenous, specific co-chaperone (middle panel) was assayed by co-immunoprecipitation. For NUDCD2, a 3xHA tagged construct was co-transfected with FBXO24 and the blot probed with an anti-HA antibody.

C) Evolution of the NUDC protein family and their client specificity. NUDC, NUDCD2 and NUDCD3 each recognize distinct β-propeller folds. NUDCD1, in contrast, associates with proteins with an unrelated fold (RNA helicases and the COPI complex), but it itself contains a β-propeller domain. The COPI complex image used with permission from Science Magazine.

D) Genomes that encode the Kelch-domain specific co-chaperone NUDCD3 contain significantly more proteins with Kelch domains than genomes without NUDCD3. The number of Kelch domains was analyzed in each of 147 fully sequenced eukaryotic proteomes. Normalized Kelch domain abundance (as a fraction of total number of proteins) was compared between species that have NUDCD3 ortholog and those that do not. Histograms display the t-statistic distribution for 10,000 random eukaryotic proteins that were assayed for similar evolutionary cooccurrence with Kelch domains. Orange line shows the t-statistic for NUDCD3 and black lines show the t-statistic for three other co-chaperones that are not Kelch-specific.

E) Evolutionary analysis of LRR domain evolution with LRR-specific co-chaperone SGT1, performed as in D. See also Figure S5.