A minority of foci or pan-nuclear apoptotic staining of gamma H2AX in the S phase after UV damage contain DNA double-strand breaks

作者：de Feraudy Sebastien^*; Revet Ingrid; Bezrookove Vladimir; Feeney Luzviminda; Cleaver James E

来源：Proceedings of the National Academy of Sciences, 2010, 107(15): 6870-6875.

DOI：10.1073/pnas.1002175107

• DNA damage response
• replication
• UV irradiation
• apoptosis
• 53BP1

UV irradiation induces histone variant H2AX phosphorylated on serine 139 (gamma H2AX) foci and highlevels of pan-nuclear gamma H2AX staining without foci, but the significance of this finding is still uncertain. We examined the formation of gamma H2AX and 53BP1 that coincide at sites of double-strand breaks (DSBs) after ionizing radiation. We compared UV irradiation and treatment with etoposide, an agent that causes DSBs during DNA replication. We found that during DNA replication, UV irradiation induced at least three classes of gamma H2AX response: a minority of gamma H2AX foci colocalizing with 53BP1 foci that represent DSBs at replication sites, a majority of gamma H2AX foci that did not colocalize with 53BP1 foci, and cells with high levels of pan-nuclear gamma H2AX without foci of either gamma H2AX or 53BP1. Ataxia-telangiectasia mutated kinase and JNK mediated the UV-induced pan-nuclear gamma H2Ax, which preceded and paralleled UV-induced S phase apoptosis. These high levels of pan-nuclear gamma H2AX were further increased by loss of the bypass polymerase Pol eta and inhibition of ataxia-telangiectasia and Rad3-related, but the levels required the presence of the damage-binding proteins of excision repair xeroderma pigmentosum complementation group A and C proteins. DSBs, therefore, represent a small variable fraction of UV-induced gamma H2AX foci dependent on repair capacity, and they are not detected within high levels of pan-nuclear gamma H2AX, a preapoptotic signal associated with ATM- and JNK-dependent apoptosis during replication. The formation of gamma H2AX foci after treatment with DNA-damaging agents cannot, therefore, be used as a direct measure of DSBs without independent corroborating evidence.

• 出版日期2010-4-13