Manipulation of Stu2 acetylation sites alters chromosome transmission fidelity.

Yeast strains for this assay contained a 125 kb artificial chromosome that expressed the URA3 gene, in addition to plasmids expressing WT-Stu2 (yRM12253), K252R (yRM12261), K252Q (yRM12262), K469R (yRM12301), K469Q (yRM12260), K870R (yRM12652), K870Q (yRM12270), 2KR (yRM12266), 2KQ (yRM12267), 3KR (yRM12269), and 3KQ (yRM12268) in a stu2-Δ1::HIS3 background. (A) To quantify the frequency at which the artificial chromosome was lost in Stu2 mutants, the cells were transferred to plates containing 5-FOA or serially diluted and transferred to -leu synthetically defined plates. Because 5-FOA is toxic to yeast possessing the URA3 gene, only colonies that have lost the 125 kb artificial chromosome should grow on 5-FOA. Colonies growing on leucine deficient plates reflect the total number of cells used in the experiment. (B) Quantitative rates of chromosome transmission fidelity reflect the total number of colonies growing on 5-FOA media relative to colonies growing on -Leu synthetic deficient media following serial dilutions. Quantitative chromosomal loss data displayed as mean ± SEM of quadruplicate plates; * p S1 Supplementary file).