Genetic interactions among mutants affecting mRNA export factors Dbp5p/Rat8p, Mex67p, Sub2p and Yra1p, and the transcription factor Bur6p and truncated Rpb2tp. (A) Synthetic lethality between bur6-ts and yra1-1 or sub2-85. Double mutants yra1-1 bur6-ts [pYRA1/URA3] (yra1-1 in the Figure), sub2-85 bur6-ts [pSUB2/URA3] (sub2-85 in the Figure), mex67-5 bur6-ts [pMEX67/URA3] (mex67-5 in the Figure) and mex67-6 bur6-ts [pMEX67/URA3] (mex67-6 in the Figure) were streaked on SC-ura (as control) or 5′-FOA (to analyze synthetic lethality) and incubated at 30 °C for 4 days. (B) rat8-2 mutant was transformed with a C-terminal deletion of RPB2 carried on the multicopy plasmid YEplac181 (YEp-rpb2t), the wild type RPB2 gene cloned in YEplac181 (YEp-RPB2) or the empty vector (YEplac181). Serial dilutions (1:10) of transformed cells were spotted onto YPD plates and incubated for 3 days at different temperatures. A yeast strain carrying the DBP5 gene under the control of the regulatable promoter containing tetO (PtetO-DBP5) was transformed with the indicated plasmids. Transformed cells were spotted onto YPD or YPD containing 10 mg/L doxycycline and incubated for 3 days at 30 °C. sub2-85 and yra1-1 mutants were transformed with the YEp-rpb2t plasmid or the empty vector YEplac181 and growth was analyzed by spotting serial dilutions of the transformants on YPD plates and incubation at different temperatures.
Estruch et al. BMC Genetics 2012 13:80 doi:10.1186/1471-2156-13-80