Double strand breaks are very dangerous lesions in DNA and must be repaired to allow cells to complete replication and transcription. Cells with deficiencies in double strand break repair are subject to genomic instability and individuals with these deficiencies are often at elevated risk of cancer. The presence of double strand breaks can be determined by examining cells for the phosphorylated form of a histone protein variant known as H2AX. Phosphorylation of the protein occurs rapidly in the vicinity of a double strand break, and persists until the break is repaired. Consequently phospho-H2AX serves a surrogate marker for breaks. This species can be easily detected by immunofluorescence techniques, and thus can be detected in single cells. We are testing the hypothesis that double strand breaks are presented in elevated levels in cells from aged individuals, as compared with younger individuals, by determining the level of phospho-H2AX in blood derived cells from donors in the BLSA program. The results indicate that cells from older individuals contain more frequent H2Ax sites than cells fromyounger individuals. The frequency We are also measuring DNA repair at the single cell level by gel electrophoresis assays.
