. 2020 Nov 20;2(1):100014. doi: 10.1016/j.xhgg.2020.100014

Table 2.

Phenotypic analyses of potentially pathogenic POLR2A variants in yeast

RPB1 alleles	POLR2A (Human)	Individual with variant	YPD 30°C	YPD 37°C	SC-Leu MPA (20μg/ml)	YPD + Formamide (3%)	SC-Lys (Spt^-)	YPRaf/Gal	SC-His
WT	WT	NA	+	+	MPA^R	Formamide^R	-	Gal^s	-
Pro24Arg	Pro28Arg	9	-/+	-	MPA^R	Formamide^S	-	Gal^s	-
Glu26Val	Glu30Val	NA	+	+	MPA^R	Formamide^R	-	Gal^s	-
Glu104His	Arg108His	10	+	+/-	MPA^R	Formamide^R	-	Gal^s	-
Glu104Leu	Arg108Leu	NA	+/-	--/+	MPA^R	Weak Formamide^S	-	Gal^s	-
Arg134Trp	Arg140Trp	11	-/+	-	MPA^R	Formamide^S	-	Gal^s	-
Arg175Trp	Arg192Trp	NA	+	+	MPA^R	Formamide^R	-	Gal^s	-
Gln313Cys	Arg327Cys	NA	+	+	MPA^R	Formamide^R	-	Gal^s	-
Leu388Val	Leu402Val	NA	+	+	MPA^R	Formamide^R	-	Gal^s	-
Asp423ΔIle424Δ	Asp437_Leu438del	NA	+	+	MPA^R	Weak Formamide^S	-	Gal^R	-
Asp423Δ	Asp437del	NA	+	+	MPA^R	Weak Formamide^S	-	Gal^R	-
Ile424Δ	Leu438del	1	+	+	MPA^R	Formamide^R	-	Gal^R	-
Ala1069Val	Ala1092Val	12	+	+	MPA^R	Formamide^R	-	Gal^R	-
Thr1272Ala	Thr1297Ala	NA	+	+	MPA^R	Formamide^R	-	Gal^s	-/+
Gly1388Arg	Gly1418Arg	6	+	+	MPA^R	Formamide^R	-	Slight Gal^R	-
Gly1388Val	Gly1418Val	NA	+	+	MPA^R	Formamide^R	-	Slight Gal^R	-

Table summarizing phenotypic analysis of potentially pathogenic POLR2A variants in yeast by spot assay (Figure 3). General (growth at standard propagation temperature on standard medium YPD 30°C) or conditional growth defects (YPD 37°C), YPD in presence of 3% formamide (a protein-folding stressor), or minimal media that enable detection of transcription-related phenotypes are shown: addition of drug mycophenolic acid (MPA) detecting defective IMD2 expression, medium lacking lysine (SC-Lys) for detection of altered transcription due to Suppressor of Ty phenotype (Spt-) at the lys2-128∂ reporter allele, rich medium YP with 2% raffinose and 1% galactose for detection of altered transcription apparent as suppression of the galactose toxicity (Gal^S) imposed by the gal10Δ56 reporter allele (YP Raf/Gal), medium lacking histidine for detection of altered transcription allowing constitutive expression of the imd2Δ::HIS3 transcriptional reporter (SC-His). Shaded cells indicate a phenotype different from WT, with darker shading indicating a stronger phenotype than lighter shading. Strength of growth is indicated as robust growth (+), reduced growth (+/−), weak growth (−/+), severe growth defect (−−/+), no growth (−).