To determine whether the physical differences between high-and low-LET radiation are reflected in the biological responses of exposed cells, we detailed phosphoprotein profiles of three proteins functional in radiation repair and signal transduction.Detailing γH2AX, pATF2 Ser 490/498 and pSMC1 Ser 957 kinetics following X-ray and ironion exposure also provides a window into understanding the underlying cellular responses. Phosphorylated forms of these proteins have been documented to co-localize at sites of double strand breaks (DSBs) after low-LET exposures, and two of these phosphorylations, pATF2 and pSMC1, are specifically ATM dependent. Flow cytometry-based methods were used to quantify total levels of each phospho-protein at various times after radiation. As expected, we observed a greater induction and persistence in γH2AX after iron-ion (high-LET) exposure as compared to X-ray (low-LET). In contrast, pATF2 and pSMC1 showed markedly lower induction levels following Fe exposure as compared to equivalent doses of X-ray. Quantification of pATF2 and pSMC1 foci revealed fewer cells containing foci, and fewer foci per cell after iron-ion compared to X-ray exposure. These findings suggest that ATM responds to DSBs induced by high-LET radiation differently from DSBs induced by low-LET radiation.2
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