Fig. 3

SpNatA ΔN-K6E does not rescue growth of ScNatAΔ cells at high temperature. a Confirming gene disruption of ARD1 and NAT1 in a ScNatAΔ strain by colony PCR using ARD1 and NAT1 primers. ARD1-specific PCR product 1024 bp (ARD1 717 bp + gene-specific sequence 307 bp). NAT1-specific PCR product 2976 bp (NAT1 2565 bp + gene-specific sequence 411 bp). b SpNatA expression was confirmed by immunoblot analyses using anti-HA (to detect HA-SpNaa15) and anti-V5 (to detect SpNaa10-V5). Anti-Zwf1 served as loading control. c Wild-type (W303-1A) and ScNatAΔ yeast cells transformed with empty pBEVY plasmid, wild-type SpNatA, or SpNatA ΔN-K6E were grown to early log-phase in SD-Ura medium. Ten-fold serial dilutions were spotted onto YPD and SD-Ura agar plates and incubated for 2 days at 30 or 38 °C. wt; wild-type