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. 2014 Apr 7;42(10):6183–6195. doi: 10.1093/nar/gku261

Figure 4.

Figure 4.

Mutation of histone H3 results in decreased expression of met-8. (A) Diagram depicting procedures used to create the homokaryotic H3K9R knock-in strain by homologous recombination. The asterisk in the H3K9R ORF indicates the location of the Lys9-to-Arg mutation at the N terminus of H3. (B) Diagram showing sequencing of the point mutation (from codon AAG to AGG) and a nonsense mutation (from codon TCC to TCG) in serine10 to introduce a SalI site in the H3K9R mutant. Sequencing result from an H3K9R strain showing that it is a homokaryotic H3K9R knock-in strain. (C) Western blot analysis of global H3 and H3K9 trimethylation in the WT, ku70RIP, cul4KO, dim-5KO and H3K9R strains. (D) Phenotypes of aerial hyphae and conidia of wild-type, ku70RIP, cul4KO, dim-5KO and H3K9R strains on plates (30°C, 32 h). (E) Growth rates of wild-type, cul4KO, dim-5KO and H3K9R strains, measured at 25°C by race tube assays in constant darkness after 1 day of light treatment. (F) Northern blot analysis showing met-8 mRNA levels in wild-type, ku70RIP and H3K9R strains.