Cuf2 affects RNA polymerase II chromatin occupancy at fzr1+ and wtf13+ genes.
pat1-114/pat1-114 cuf2+/cuf2+ and pat1-114/pat1-114 cuf2Δ/cuf2Δ strains were synchronously induced into meiosis. At the 6-h meiotic time point, chromatin was immunoprecipitated using RNA polymerase II CTD-specific antibodies. Quantification of immunoprecipitated chromatin was performed by qPCR using primers that allowed amplification of several specific DNA regions (A, C, D, ORF A, ORF B, ORF C, and 3’UTR A), which spanned fzr1+ (panel A), wtf13+ (panel B), and mei4+ (panel C) genes. ChIP results are presented as enrichments of specific noncoding and coding regions (fzr1+, wtf13+, or mei4+) relative to a 18S ribosomal DNA coding region. Data were calculated as values of the largest amount of chromatin measured (fold enrichment). Results are shown as the averages ± S.D. of a minimum of three independent experiments. Arrows indicate the direction of transcription. Nucleotide numbers refer to the position relative to the A of the initiator codon of each gene (fzr1+, wtf13+, or mei4+).