Table 1.

Summary of DNA-PKcs autophosphorylation and catalytic mutants

DNA-PKcs constructSubstitution site(s)Level of:Description (reference)
RadioresistanceKinase activity
Wild typeFull-length human DNA-PKcs++++++++++(36)
VectorpCMV6(36)
ABCDE>AlaT2609A, S2612A, T2620A, S2624A, T2638A, T2647A− − −a+++++Limited end processing; substantially more radiosensitive than V3 parental strain (12)
ABCDE>AspT2609D, S2612D, T2620D, S2624D, T2638D, T2647D±+++++Normal end processing (12)
PQR>AlaS2023A, S2029A, S2041A, S2053A, S2056A++++++++Excessive end processing (10)
PQR>AspS2023D, S2029D, S2041D, S2053D, S2056D++++++++++Limited end processing; poorly expressed (10)
M>AlaS3205A++++++++++ATM site in vivo; Fig. 4 (16)
M>AspS3205D++++++++++ATM site in vivo; Fig. 4 (16)
T>AlaT3950A++++++++Putative T-loop site (14)
T>AspT3950DPutative T-loop site (14)
JK>AlaT946A, S1004A++++++++Highly conserved S/T-Q; Fig. 5
JK>AspT946D, S1004D+++++Highly conserved S/T-Q; Fig. 5
N>AlaS56A, S72A++++++++++Highly conserved, N-terminal S/T hydrophobic; Fig. 6
N>AspS56D, S72D±Highly conserved, N-terminal S/T hydrophobic; Fig. 6
L>AlaT1865A++++++++Highly conserved S/T-Q; see Fig. S2 in the supplemental material
L>AspT1865D++++++++Highly conserved S/T-Q; see Fig. S2
LZ>AlaS1470A, S1546A++++++++++Highly conserved S/T-Q within LRR; see Fig. S2
LZ>AspS1470D, S1546D++++++++++Highly conserved S/T-Q within LRR; see Fig. S2
Lys>ArgK3752RConserved K in kinase domain (22)
Asp>AlaD3921APutative catalytic aspartate
  • a −−−, increased radiosensitivity compared to that of vector-only controls.