Skip to main content
. Author manuscript; available in PMC: 2023 Sep 29.
Published in final edited form as: Cell. 2022 Sep 15;185(20):3671–3688.e23. doi: 10.1016/j.cell.2022.08.014

Figure. 5. MMEJ by WT and mutant G2L4 and GII RTs.

Figure. 5.

(A and B) Biochemical assays. Reactions were done with pre-annealed double-stranded DNAs having 3’ overhangs ending with complementary CCGG-3’ (panel A) or non-complementary CCAA-3’ (panel B) sequences. The pre-annealed oligonucleotides are denoted D1/D2 (left side) and D1’/D2’ (right side), with D1/D1’ 5’-labeled (red star) and D2/D2’ 3’-blocked. The numbers to the left of the gels indicate the positions of size markers in a parallel lane. The plots show the fraction of substrate converted to products running between the 100- and 150-nt size markers. Tables show rate constants (kobs) and amplitudes (Ampl.) obtained by fitting the data to a first-order rate equation. Values in parentheses indicate that the amplitude was fixed at the given value because the reaction did not reach an end point during the experiment.

(C) High-throughput sequencing of MMEJ products with DNA substrates having CCGG-3’ microhomologies. Schematics of different products are shown to the left. Red and green indicate nucleotides at the 3’ end of nascent DNA or added by terminal transferase (TT) to the 3’ end of the nascent DNA, respectively. The stacked bar graphs show the proportions of different products in different samples.