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Genetic Control of Lithium Sensitivity and Regulation of Inositol Biosynthetic Genes

Figure 4

PO regulates gene expression.

A, Expression of ino1 in wild type, dpoA null cells and wild type cells following overnight treatment with 1.2 mM Z-pro-L-prolinal (PO inhibitor), measured by Northern analysis. rnlA expression was used as a loading control. The graph shows the expression of ino1 quantified on a phosphoimager (Biorad) and normalised to rnlA relative to wild type or DMSO carrier treated controls (mean ± standard error of 3 independent experiments). B, The expression of genes involved in inositol metabolism (Table S1). Gene expression in wild type cells following overnight treatment with Z-pro-L-prolinal or the equivalent amount of DMSO carrier, and dpoA null mutant cells was measured by qRT-PCR, and normalised to rnlA expression levels. Values plotted are relative to the carrier control cells (mean ± standard error of 3 independent experiments). * P<0.05, ** P<0.01, *** P<0.005, T-Test. C, mipp1 is required for the regulation of gene expression by PO. Expression of ino1 and impA1 in wild type and mipp1 null cells following overnight treatment with Z-pro-L-prolinal or the equivalent amount of DMSO carrier, was measured by qRT-PCR, and normalised to rnlA expression levels. Values plotted are relative to the carrier control wild type cells (mean ± standard error of at least 3 independent experiments). There is a significant difference in gene expression between wild type and mipp1 null mutant following PO inhibition (** P<0.01, T-Test). D, Expression levels of ino1, impA1 and mipp1 in wild type, mipp1 null mutant and cells over-expressing Mipp1, IpkA1 or IpkB. Expression was calculated relative to either wild type, or wild type cells transformed with the empty expression vector as appropriate. All samples were normalised to rnlA as a loading control and the values plotted are the mean ± standard error of 3 independent experiments (*P<0.05, ** P<0.01 T-test). ND = not determined.

Figure 4

doi: https://doi.org/10.1371/journal.pone.0011151.g004