Table 2.

Phenotypic analysis of RRM1 mutations

Strain YC504 containing the YC360 plasmid (PAB1-URA3) instead of theYC504 plasmid (Flag-PAB1-TRP1) was transformed with each Flag-PAB1-TRP1 variants as described in the Table. The ability to subsequently lose the YC360 plasmid was assayed on 5-fluoroorotic acid-containing plates (Yao et al 2007). Transformed cells incapable of losing YC360 were considered displaying a synthetic lethality with caf1. cdc33-1 carries mutations in elF4E rendering it unable to grow at 37°C under glucose growth conditions (Lee et al 2010). Growth at 41°C glucose and 34°C glycerol was monitored for cells containing only the RRM1 variant as indicated following loss of the YC360 plasmid. All other RRM1 alterations listed in Table 1 elicited no discernable phenotypic effects. ‘++’ indicates good growth and ‘−‘ indicates no growth.

RRM1 mutation	Growth, 41°C glucose	Growth in cafl background, 30°C glucose	Growth in cdc33-1 background, 34°C glycerol
PAB1-wt	++	++	++
PAB1-Y83V	++	−	−
PAB1 Y41A	++	−	−
PAB1-R110A	++	−	++
PAB1-C70S C109V	−	++	++
PAB1-T102D P103V	++	++	++
PAB1-RRM1	−	−	−