Table 5.
DNA sequence analysis of descendants of a gap-lesion plasmid carrying a sitespecific TT 6-4 PP after repair in polη-proficient and polη-deficient human cells
| DNA sequence opposite TT CPD | Primary fibroblasts | SV40-transformed fibroblasts | Burkitt's lymphoma cells | ||||
|---|---|---|---|---|---|---|---|
| POLH+/+ | POLH−/− | POLH+/+ | POLH−/− | POLH+/+ | POLH−/− | ||
| Number of isolates (%) | |||||||
| C-AA-C | 59 (46) | 40 (40) | 43 (47) | 22 (34) | 86 (68) | 54 (55) | |
| A-AA-C | - | - | 1 (1) | - | 2 (2) | - | |
| T-AA-C | 1 (1) | 1 (1) | 7 (8) | 1 (2) | - | - | |
| C-CA-C | 1 (1) | - | 1 (1) | 2 (3) | - | 1 (1) | |
| C-GA-C | 5 (4) | 5 (5) | 2 (2) | 2 (3) | 2 (2 ) | 1 (1) | |
| C-TA-C | 2 (2) | 3 (3) | 1 (1) | 2 (3) | - | 2 (2) | |
| C-AC-C | 2 (2) | 1 (1) | 3 (3) | 2 (3) | 1 (1) | - | |
| C-AG-C | 3 (2) | - | - | 2 (3) | 3 (2) | 1 (1) | |
| C-AT-C | 3 (2) | - | 3 (3) | 1 (2) | 1 (1) | 1 (1) | |
| C-AA-A | 25 (20) | 32 (32) | 22 (24) | 17 (27) | 11 (9) | 15 (15) | |
| C-AA-G | - | 1 (1) | - | - | - | - | |
| C-AA-T | - | - | - | 1 (2) | - | 1 (1) | |
| C-ΔA-C | - | - | 4 (4) | 1 (2) | 1 (1) | 3 (3) | |
| C-AΔ-C | 1 (1) | - | - | - | 5 (4) | 8 (8) | |
| C-AA-Δ | - | - | - | 1 (2) | - | - | |
| C-AΔ-Δ | - | - | - | - | - | 1 (1) | |
| A-CA-C | - | 1 (1) | - | - | - | - | |
| A-GA-C | - | - | - | 1 (2) | - | - | |
| A-TA-C | 1 (1) | - | - | - | - | - | |
| T-CA-C | 1 (1) | - | - | - | - | - | |
| C-CC-C | - | 1 (1) | - | - | - | - | |
| C-GC-C | 1 (1) | - | - | - | - | - | |
| C-TG-C | 1 (1) | - | - | - | 1 (1) | - | |
| C-TT-C | - | 2 (2) | - | 1 (2 ) | 1 (1) | 1 (1) | |
| A-AG-C | - | - | 1 (1) | - | - | - | |
| C-GA-A | 3 (2) | 2 (2) | - | 4 (6) | 4 (3) | 2 (2) | |
| C-TA-A | 7 (5) | - | - | 2 (3) | 2 (2) | - | |
| Non-TLS events | 12 (9) | 10 (10) | 3 (3) | 2 (3) | 6 (5) | 8 (8) | |
| Total | 128 (100) | 99 (100) | 91 (100) | 64 (100) | 126 (100) | 99 (100) | |
Plasmids were extracted from kanR colonies obtained in the experiments described in Table 4, and subjected to DNA sequence analysis. The sequence opposite the site of the TT 6-4 PP is shown in the 5' to 3' direction. Accurate TLS is represented by the sequence 5'-CAAC-3'. The underlined nucleotides are those located opposite the original TT 6-4 PP. Mutations are presented by bold type. Δ represents a one-nucleotide deletion. Non-TLS events include big deletions and insertion of sequences from the control plasmid.