Table 1.

Probing the Role of PolD1 and PolD2 in NHEJ of 5’-Overhang DSBs In Vivo

Strain	NHEJ Efficiency (%)	NHEJ Fidelity (%)	Junctions Sequenced	Deletions	Templated Fill-ins
Wild-type	100	33	22*	8*	21*
ligD-(D136A- D138A)	33	74	10	10	7
ΔpolD2	153	36
ΔpolD1	117	22
ligD-(D136A- D138A) ΔpolD1	21	72
ligD-(D136A- D138A) ΔpolD2	49	70
ligD-(D136A- D138A) ΔpolD1 ΔpolD2	12	71	15	13	11

The efficiencies of linear transformation with the Asp7181-cut reporter plasmid are expressed relative to wild-type (defined as 100%). Fidelity is calculated as [blue colonies/(blue+white colonies)] × 100. Each value for efficiency and fidelity is the mean of three independent plasmid transfections and is representative of 2–3 independent experiments. Using primers upstream and downstream of the repair junctions, the repair junctions for non-faithful repair events (white colonies) were amplified by colony PCR. The junctions were sequenced and the repair events were characterized. The number of events with terminal deletions and templated additions are indicated.

^*The junction sequence data from wild type M. smegmatis is reproduced from ref. 21.