Nature of mutation in the human hprt gene following in vivo exposure to ionizing radiation of cesium-137.

Abstract

The current study comprises the analysis of mutations in 10 individuals accidentally exposed to cesium-137 during the 1987 radiological accident in Goiânia, Brazil. Their exposures were among the highest experienced, ranging from 1 to 7 Gy. Peripheral T-lymphocyte samples were obtained 3.3 years after the original exposure and mutation was studied at the hprt locus using the 6-thioguanine-resistance selection assay. The mutational spectrum for the exposed population is comprised of 90 independent mutants. Based on T-cell receptor analysis, only 5% (5/95) were clonally related. Mutants were initially studied using RT-PCR and directly sequenced using an automated laser fluorescent DNA sequencer. Mutants that repeatedly failed to produce cDNAs were studied using a multiplex PCR assay with genomic DNA. Missense mutations were the most frequent event recovered, comprising 40% (23/57) of the spectral sample. An excess of events involving A:T base pairs was observed, exhibiting a significant difference (chi 2 = 12.7, P = 0.0004) when compared to the spontaneous spectrum. This finding may reflect the effect of ionizing radiation-induced damage, suggesting a potential similarity to radiation effects in prokaryotes. At the genomic level, 36.7% (33/90) of the mutants exhibited gross structural alterations, as detected by multiplex PCR. Deletion events were over-represented in our spectral sample, displaying a twofold increase when compared to the frequency observed in the spontaneous mutation database.