FIGURE 2.

Bud23 promotes the recruitment of the Rps0-cluster proteins to 40S precursors. (A, left) Ectopic expression of RPS2 suppressed the growth defect of bud23Δ as shown by 10-fold serial dilutions of wild-type (BY4741) or bud23Δ (AJY2676) cells transformed with an empty vector (pRS415) or a vector encoding BUD23 (pAJ2154) or RPS2 (pAJ2960) spotted on SD Leu- media and grown for 2 d at 30°C. (Right) Ectopic expression of RPS2 suppressed the growth defect of bud23Δ, as shown by the quantification of doubling time in minutes of the same strains used in A. See Materials and Methods for details. Data are shown as mean doubling time ± standard deviation of biological duplicates. Significance was calculated by two-way analysis of variance (ANOVA) with Sidak correction for multiple comparisons using GraphPad Prism 9 for Mac iOS (www.graphpad.com) (adjusted P = 0.9155 [ns]; P = 0.0035 [**]). (B,C) Bud23-depletion decreased the association of Rps0, Rps2, and Rps21-HA as shown by western blotting of specific factors associated with affinity-purified particles using Enp1-TAP as bait. Strains AJY4748 (Bud23+) and AJY4750 (Bud23−) that were transformed with pAJ5124 for B and AJY4749 (Bud23+) and AJY4751 (Bud23−) for C were each cultured in SD Ura- or YPD to early exponential phase, then treated with 0.5 mM auxin for 10 min prior to collection.