Preferential repair of ionizing radiation-induced damage in the transcribed strand of an active human gene is defective in Cockayne syndrome.
Cells from patients with Cockayne syndrome (CS), which are sensitive to killing by UV although overall damage removal appears normal, are specifically defective in repair of UV damage in actively transcribed genes. Because several CS strains display cross-sensitivity to killing by ionizing radiation, we examined whether ionizing radiation-induced damage in active genes is preferentially repaired by normal cells and whether the radiosensitivity of CS cells can be explained by a defect in this process. We found that ionizing radiation-induced damage was repaired more rapidly in the transcriptionally active metallothionein IIA (MTIIA) gene than in the inactive MTIIB gene or in the genome overall in normal cells as a result of faster repair on the transcribed strand of MTIIA. Cells of the radiosensitive CS strain CS1AN are completely defective in this strand-selective repair of ionizing radiation-induced damage, although their overall repair rate appears normal. CS3BE cells, which are intermediate in radiosensitivity, do exhibit more rapid repair of the transcribed strand but at a reduced rate compared to normal cells. Xeroderma pigmentosum complementation group A cells, which are hypersensitive to UV light because of a defect in the nucleotide excision repair pathway but do not show increased sensitivity to ionizing radiation, preferentially repair ionizing radiation-induced damage on the transcribed strand of MTIIA. Thus, the ability to rapidly repair ionizing radiation-induced damage in actively transcribing genes correlates with cell survival. Our results extend the generality of preferential repair in active genes to include damage other than bulky lesions.
Leadon
S A
SA
Department of Radiation Oncology, University of North Carolina, Chapel Hill 27599-7512.
Cooper
P K
PK
D015139:Blotting, Southern; D002460:Cell Line; D003057:Cockayne Syndrome / Q000235:genetics*; D004247:DNA / Q000528:radiation effects*; D004249:DNA Damage / Q000528:radiation effects*; D004260:DNA Repair; D015870:Gene Expression; D005796:Genes; D006801:Humans; D066298:In Vitro Techniques; D008668:Metallothionein / Q000235:genetics; D014965:X-Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics
D004247:DNA; D008668:Metallothionein