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. Author manuscript; available in PMC: 2011 Feb 1.
Published in final edited form as: Fungal Genet Biol. 2009 Sep 13;47(2):107. doi: 10.1016/j.fgb.2009.09.002

Figure 5. Overexpression of CaRCN1 rescues the viability of pmc1 strains.

Figure 5

The C. albicans or S. cerevisiae RCN1 gene was cloned into plasmid pYES for expression in S. cerevisiae under the control of the GAL1 promoter. The plasmids were sequence confirmed and then strains pmc1::TRP1 vcx1 Δ (K665), pmc1::TRP1 (K605), and vcx1 (K661) were transformed. Under conditions of high calcium, calcineurin promotes the expression of PMC1 and inhibits Vcx1 function. pmc1 strains are unable to grow at high calcium levels unless calcineurin inhibition of Vcx1 is abolished. Overexpression of either the CaRCN1 or ScRCN1 gene allows growth of pmc1 strains in high calcium suggesting that calcineurin function is inhibited, whereas presence of the vector alone did not effect the growth of pmc1 strains. Inhibition of calcineurin also prevents the strong upregulation of PMC1 in response to high calcium, thus the high calcium conditions were chosen so that the basal expression of PMC1 was sufficient to support growth when calcineurin was inhibited in the vcx1 background. Strains were plated on solid YPD medium alone or containing 0.3 M CaCl2. The carbon source was either 2% glucose or 2% galactose, as indicated.