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Fig 3
hsdS alleles downstream and upstream of xerD are expressed and methylate the genome. (A) Expression of SsuCC20p genes was verified by PCR on reverse-transcribed RNA, using gene-specific qPCR primers. PCR products were run on a 1% agarose gel. RT + or − indicates the presence of reverse transcriptase in the reverse transcription reaction; marker lanes were loaded with the 1 kb+ ladder. (B) PCR amplification of xerD-hsdS E fragment from genomic DNA (gDNA) and cDNA. RT + or − indicates the presence of reverse transcriptase in the reverse transcription reaction; marker lanes were loaded with the 1 kb+ ladder. (C) 861160 WT methylome; m6A methylated sites in genome are indicated on the genome with the color corresponding to the DNA motif; gray indicates sites that have either of the two DNA motif sites. (D) Homology of SsuCC20p TRDs to other type I RM system hsdS genes; identified DNA target sequences are indicated between brackets; for details, see Fig. S2. (E) Proposed target DNA motif of HsdS A and HsdS E.