of nuclear foci in response to DNA DSBs differs from the formation of the “apoptotic γH2AX ring” (Solier and Pommier, 2009). They demonstrated that γH2AX ring staining is an early apoptosis indicator that precedes a global nuclear staining or pan-nuclear staining and apoptotic body formation. The main driver of this particular phosphorylation is DNA-PK in contrast to ATM and ATR associated with γH2AX nuclear focus formation. This morphology variation could potentially be used to discriminate DNA DSBs from other forms of DNA damage. γH2AX could also act as a cell cycle checkpoint (Downey and Durocher, 2006). H2AX could become phosphorylated at any point during the cell cycle, including during mitosis while other DDR proteins are limited to interphase cells (Nakamura et al., 2010). It has been suggested that DSB repair mechanisms may be suspended during mitosis. However, γH2AX foci continue
to form during mitosis. The foci act Ku-0059436 mouse as indicators to activate the repair mechanisms as soon as the cell has finished the division process. If the DNA DSB occurs in G1, the cell Epacadostat price cycle would stop to prevent the cell moving into S-phase with damaged DNA. Likewise, DNA replication could be slowed if the DNA DSB has occurred in S-phase, so that the repair mechanisms could act before the DNA polymerase reaches the damaged section. Finally, when the damage occurs in G2-phase, the cell is prevented from moving into mitosis, avoiding the fracture of chromosomes during anaphase and cytokinesis (Jackson, 2002). Following the induction of DSBs, phosphorylation of the serine 139 residue starts within minutes, reaching a plateau at around 30 min after damage occurs (Paull et al., 2000). The phosphorylation then decreases over a period of hours (Rogakou et al., 1998). The mechanism of γH2AX elimination has not been fully unravelled. There are multiple phosphatases involved in γH2AX dephosphorylation. Dephosphorylation could occur directly on the chromatin or could happen after the histone has been displaced from the nucleosomes (Chowdhury et al., 2005 and Redon et al., 2011a). Both mechanisms could potentially occur simultaneously, independent
of the location of the γH2AX in the foci. Other mechanisms mentioned by Bao involve histone chaperone proteins in the process of γH2AX elimination (Bao, 2011). Experiments carried out by Thalidomide Keogh and colleagues suggest that the loss of γH2AX could be triggered not only by DSB repair but also by the activation of steps that precede DSB repair (Keogh et al., 2006). However, some of their results seem to indicate that γH2AX loss is not mediated by single-stranded DNA resection, one of the cellular responses to DSBs. There are several reasons why γH2AX is used to detect DSBs. The formation of γH2AX is proportional to the amount of DSBs, giving a direct 1:1 correlation to existing damage (Sedelnikova et al., 2002). This correlation indicates that for every DSB one nuclear focus would be created.