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The Origin Recognition Complex Interacts with a Subset of Metabolic Genes Tightly Linked to Origins of Replication

Figure 5

Transcriptional activity promoted ORC binding to an ORF.

(A) Histograms of expression levels of the ORF–ORC set and of all genomic ORFs are shown. Dubious ORFs were not included in this analysis. (B) TDH3 promoter was replaced by the GAL1 promoter (pGAL1). pGAL1-TDH3 and wild-type strains were grown in glucose or galactose, RNA was isolated, and gene expression measured by reverse transcriptase PCR. Growth in glucose repressed TDH3 transcription in the pGAL1-TDH3 strain but not in an isogenic wild-type strain, whereas wild-type TDH3 was virtually unaffected by change of carbon source. As expected, expression of the GAL1 gene was repressed in glucose and induced in galactose. ENO2 expression was monitored as a control. (C) Results of directed ORC ChIPs on the wild-type and pGAL1-TDH3 strains grown in glucose and galactose are shown. Averages of two to four independent biological replicates are plotted for each condition, with error bars representing one standard deviation. Repression of pGAL1-TDH3 transcription by glucose reduced ORC binding to TDH3 ORF, while induction of GAL1 in galactose increased ORC binding there by 2.5-fold.

Figure 5

doi: https://doi.org/10.1371/journal.pgen.1000755.g005