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PMID:15084308
| Citation |
Bannister, LA, Waldman, BC and Waldman, AS (2004) Modulation of error-prone double-strand break repair in mammalian chromosomes by DNA mismatch repair protein Mlh1. DNA Repair (Amst.) 3:465-74 |
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| Abstract |
We assayed error-prone double-strand break (DSB) repair in wild-type and isogenic Mlh1-null mouse embryonic fibroblasts containing a stably integrated DSB repair substrate. The substrate contained a thymidine kinase (tk) gene fused to a neomycin-resistance (neo) gene; the tk-neo fusion gene was disrupted in the tk portion by a 22bp oligonucleotide containing the 18 bp recognition site for endonuclease I-SceI. Following DSB-induction by transient expression of I-SceI endonuclease, cells that repaired the DSB by error-prone nonhomologous end-joining (NHEJ) and restored the correct reading frame to the tk-neo fusion gene were recovered by selecting for G418-resistant clones. The number of G418-resistant clones induced by I-SceI expression did not differ significantly between wild-type and Mlh1-deficient cells. While most DSB repair events were consistent with simple NHEJ in both wild-type and Mlh1-deficient cells, complex repair events were more common in wild-type cells. Furthermore, genomic deletions associated with NHEJ events were strikingly larger in wild-type versus Mlh1-deficient cells. Additional experiments revealed that the stable transfection efficiency of Mlh1-null cells is higher than that of wild-type cells. Collectively, our results suggest that Mlh1 modulates error-prone NHEJ by inhibiting the annealing of DNA ends containing noncomplementary base pairs or by promoting the annealing of microhomologies. |
| Links |
PubMed Online version:10.1016/j.dnarep.2004.01.001 |
| Keywords |
Adaptor Proteins, Signal Transducing; Animals; Base Pair Mismatch/genetics; Base Sequence; Carrier Proteins; Cell Line; Chromosome Breakage/genetics; Chromosomes, Mammalian/genetics; Chromosomes, Mammalian/metabolism; DNA Repair; Gene Deletion; Mice; Mice, Knockout; Neoplasm Proteins/deficiency; Neoplasm Proteins/genetics; Neoplasm Proteins/metabolism; Nuclear Proteins |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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See also
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