Figure 4

Tpk binding to EFG1 and MSB2 loci. A. Binding to EFG1. Coordinates of EFG1 and neighbouring genes on chromosome R are shown on top. Round circles (dotted lines) indicate the position of Efg1 binding during growth in the yeast form [13]; the kinked arrow indicates the transcript start site. The genomic localization of Tpk1 and Tpk2 was determined by ChIP chip assays on strains containing HA-tagged Tpk proteins relative to an unmarked control strain. For Tpk1 localization, the strain pair II (TPK1/tpk1)/AF1004 (TPK1-3× HA/tpk1) and for Tpk2 localization, the strain pair TPK7 (TPK2/tpk2)/AF1005 (TPK2-3× HA/tpk2) was used in ChIP chip. Graphs represent duplicates of Tpk^HA occupancy at the EFG1 locus, ordinates represent scaled log₂ ratios. B. Binding to MSB2. Tpk2 enrichment in (a) the MSB2 ORF of yeast cells or (b) in the MSB2/ORF19.1488 upstream region of cells following hyphal induction; the most significant binding sequences are indicated by the dotted box. Transcript levels of MSB2 (c) or ORF19.1488 (d) in yeast or in induced cells were determined by qPCR in wild-type CAF2-1 (TPK2) or in tpk2Δ mutant cells (strain TPO7.4) growing in the yeast form or induced 30 min for hypha formation. Transcript levels relative to the ACT1 transcript (RTL) are shown for two biological replicates (black and grey bars) indicating means and standard deviations for three technical replicates.