Genome-wide Analysis of Gene Expression Regulated by the Calcineurin/Crz1p Signaling Pathway in Saccharomyces cerevisiae

In Saccharomyces cerevisiae , the Ca 2+ /calmodulin-dependent protein phosphatase, calcineurin, is activated by specific environmental conditions, including exposure to Ca 2+ and Na + , and induces gene expression by regulating the Crz1p/Tcn1p transcription factor. We used DNA microarrays to perform a comprehensive analysis of calcineurin/Crz1p-dependent gene expression following addition of Ca 2+ (200 m m ) or Na + (0.8 m ) to yeast. 163 genes exhibited increased expression that was reduced 50% or more by calcineurin inhibition. These calcineurin-dependent genes function in signaling pathways, ion/small molecule transport, cell wall maintenance, and vesicular transport, and include many open reading frames of previously unknown function. Three distinct gene classes were defined as follows: 28 genes displayed calcineurin-dependent induction in response to Ca 2+ and Na + , 125 showed calcineurin-dependent expression following Ca 2+ but not Na + addition, and 10 were regulated by calcineurin in response to Na + but not Ca 2+ . Analysis of crz1Δ cells established Crz1p as the major effector of calcineurin-regulated gene expression in yeast. We identified the Crz1p-binding site as 5′-GNGGC(G/T)CA-3′ by in vitro site selection. A similar sequence, 5′-GAGGCTG-3′, was identified as a common sequence motif in the upstream regions of calcineurin/ Crz1p-dependent genes. This finding is consistent with direct regulation of these genes by Crz1p.