Genome-wide mapping of 5´-ends and ribonucleotides in mtDNA.
(A) Circos figure of the H-strand (upper panel) and L-strand (lower panel) using 5´-End-seq and HydEn-seq. The peaks are normalized to per million reads and the maximum peak is adjusted to the maximum number of reads in the HydEn-seq library. (B) Relative amount of raw reads on the H-strand (H) or L-strand (L) in mtDNA and in the two nuclear DNA strands (reverse strand (RV) and forward strand (FW). Reads from ribonucleotide incorporations in upper panel and reads from HincII sites in lower panel. (C) Quantification of ribonucleotides in H-strand and L-strand per mtDNA molecule. (D) Southern Blot visualizing the ribonucleotide incorporation in vivo in mtDNA from HeLa-cells. Linearized mtDNA was subjected to salt (KCl) or alkali (KOH). Lanes 1 and 2, probes detecting both the H-strand and L-strand; lanes 3 and 4, probe detecting only the H-strand; lanes 5 and 6, probe detecting only the L-strand.