Figure 1.

RCN-YFP fusions supply regulatory A subunit function in yeast. RCN1-YFP and YFP-RCN1 fusions were expressed under control of the constitutive alcohol dehydrogenase promoter (Ammerer, 1983) in tpd3-1 yeast cells (van Zyl et al., 1992). A, Transformants carrying RCN-YFP fusions, an RCN1 construct, or the empty vector were streaked on duplicate YPD plates and incubated at 30°C or 37°C. The diagram at left indicates the construct carried by cells in the corresponding sectors on both plates. B, Cells carrying a native RCN1 construct, YFP-RCN1, RCN1-YFP, or a SUP35:GFP fusion (Satpute-Krishnan and Serio, 2005) were grown to early log phase and mounted for differential interference contrast (bottom panels) or fluorescence (top panels) microscopy. C, Protein extracts of cells carrying the constructs indicated were subjected to SDS-PAGE and immunoblotting, using anti-GFP antibodies to detect the fusion proteins. The positions of the RCN1-YFP (black arrows) and SUP35:GFP (asterisk) proteins are indicated at right. D, Cells carrying the constructs indicated were grown in liquid culture and 10-fold serial dilutions were spotted on plates containing YPD medium or YPD plus 600 mm NaCl. YPD plates were incubated at 30°C or 37°C and YPD NaCl plates were incubated at 30°C.