Yeast and Bacteria Growth Assay

Yeast cells were routinely grown at 30 °C in YPD (1% yeast extract, 2% peptone, 2% glucose) medium or in SD (0.15% yeast nitrogen base, 0.5% ammonium sulphate, 2% glucose) broth supplemented with the appropriate amino acids and bases as nutritional requirements [73 ]. All solid media contained 2% agar. Yeast was transformed using the lithium acetate method [74 (link)]. Bacteria were routinely grown in LB (0.5% yeast extract, 1% tryptone, 0.5% NaCl) broth with or without agar, in the presence of 100 µg/mL ampicillin, when required.
To test the in vivo function of the different truncated dbp7 alleles, a dbp7∆ strain harbouring plasmid YCplac33-DBP7 (CEN URA3 DBP7) was transformed with YCplac22 (CEN TRP1) plasmids carrying various dbp7 truncated alleles (see Table S2). Trp⁺ transformants were selected and streaked out on plates containing 5-fluoroorotic acid (5-FOA) to counter-select for the URA3 plasmid [75 (link)]. Clones growing on these plates were recovered on fresh SD-Trp plates, and their growth was then assessed on YPD and selective SD-Trp media.