Figure 6.

MMEJ-mediated intermolecular ligation in NHEJ-deficient cells. (A) Exponentially growing cells were cotransformed with ura4⁺ DNA and hDNA flanked by either microhomologous (8 bp) or homologous (205 and 401 bp) sequences to 5′- and 3′-ends of ura4⁺ DNA. (B) Number of Ura⁺ colonies obtained after independent transformations of wild-type (PN559), rhp51Δ (PN2490), rad50Δ (KT120), lig4Δ (AD459), pku70Δ (PN3773), and lig4Δpku70Δ (AD462) cells with 2 μg ura4⁺ DNA and either 2 μg microhomologous hDNA or 2 μg homologous hDNA. (C) Insertion frequency of both hDNA types. (D) Repair pathway for hDNA insertion in ura4⁺ circles based on overlapping sequences at junctions. For cotransformation with microhomologous hDNA, insertions were classified under MMEJ if the 8 bp overlapped at junctions. The number of sequenced inserts is shown in parentheses. WT, wild type.