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Gene duplication and co-evolution of G1/S transcription factor specificity in fungi are essential for optimizing cell fitness

Fig 6

Chimeric Swi4 TFs bind MCB-like ancestral motifs, but not SCBs, which are required for binding and regulation by wt Swi4.

(A-B) anti-swi4 Chromatin Immunoprecipitation (ChIP). Binding to CLN2, PCL1 and PRY2, SCB motifs containing promoters (SCB), and NRM1, MCD1, and ELO1, MCB-like motifs containing promoters (MCB-like) was analyzed by ChIP. Binding was assessed in alpha factor arrested cultures (0 min in top panel) and arrested (0 min) and released cultures (30 min and 60 min, bottom panel). Enrichment is indicated as percentage of input of Whole Cell Extract (% of WCE). ChIP analysis was carried out in (A-B) wild type (wt) and swi4Δ cells and wild type cells harboring mutations in the PRY2 promoter disrupting the core SCB motif (wt AA PRY2) and changing it to MCB-like motifs (wt MCB PRY2). (C) Analysis of PRY2 expression in wild type (wt), swi4Δ and PRY2 promoter mutants (AA and MCB-like). (D) ChIP analysis of wild type (wt) and all chimeric TFs used in the RNA seq experiment, K. lactis (KlMbp1BD-Swi4AD), C. albicans (CaMbp1BD-Swi4AD), N. crassa (NcResBD-Swi4AD) and S. pombe chimeras (SpMbp1BD-Swi4AD) for indicated promoters.

Fig 6