Damage to cells elicits increases and decreases in the expression of many genes. Recent microarray analysis has shown that these changes can involve hundreds of genes and that different stresses can invoke both a common set of genes and genes that are peculiar to particular kinds of stress (Amundson and others 1999a, 1999b). Despite the large number of affected genes, none appears to be directly involved in repair of DSBs (Tusher and others 2001). Central to most damage responses is stabilization of the tumor-suppressor gene p53, which occurs as a result of posttranslational phosphorylation or acetylation of the protein (Blattner and others 1999; Figure 1-11). Multiple potential serine and threonine residues in p53 are capable of being phosphorylated by different kinases in response to cellular stress, and several thousand combinations of modifications are possible in an irradiated cell. Resolving the functional role of any particular site can be difficult (Blattner and others 1999). The kinases include ATM, ATR, Chk1, Chk2, DNA-dependent protein kinase, and casein kinase I and II (Blattner and others 1999; Chehab and others 2000). (For the role that p53, pRb, cdc25C, chk1, chk2, 14-3-3 proteins, bub1, and the various cyclins and cyclin-dependent kinases play in radiation-induced checkpoints in G1, G2, and mitosis, see Little 1994; Jacks and Weinberg 1998; Lengauer and others 1998; Schmidt-Kastner and others 1998; Chan and others 1999; Ford and Pardee 1999; White and Prives 1999).
ATM is a centrally important kinase for X-ray damage that is activated by DNA DSBs (Bakkenist and Kastan 2003; Figure 1-11). In X-irradiated cells, phosphorylation of serine 15 and 37 interferes with the association of p53 with another protein mdm2 that also becomes phosphorylated and normally causes degradation of p53, extending its lifetime. The increased stability of p53 in irradiated cells permits it to form a tetramer and then act as a transactivating factor, increasing the expression of many other genes. Clearly, this will result in large-scale alterations of the gene expression pattern of irradiated cells that can influence their behavior. One downstream target for p53 is the cell cycle regulator protein p21; increased transcription of p21 due to p53 results in delays in the onset of DNA synthesis (the G1 checkpoint) and reduced DNA synthesis due to p21 binding the replication factor PCNA. The major response of cells to ionizing radiation is a reduction in initiation of the S phase and of replication origins during S. Another important radiation-responsive gene is GADD45; both this and p21 showed a linear dose-response relation for induction from 20 to 500 mGy with no indication of a threshold (Amundson and others 1999b).
Most of the members of the signal transduction pathways including ATM, p53, Chk1, Chk2, Brca1, and hMre11/hRad50/Nbs1 are protein products of tumor-suppressor genes. Loss of function of these members can result in genomic instability and in some instances may contribute to a series of events resulting in malignancy. They influence cell cycle checkpoints, DNA replication, DNA repair, and recombination. Thus, it is possible for a single DNA DSB to activate ATM and p53 and create a cell-wide response through this cascade of protein modifications and alterations in gene expression.
These signal transduction pathways are also activated by extracellular signals working through specific receptors on the cell membrane that then activates kinases, such as MAPKs, which phosphorylate p53. Irradiated cells also generate extracellular signals that resemble cytokines released during normal in vivo cell-cell communication processes (Herrlich and others 1992). These can, through receptors on adjacent cells or gap junctions, result in activation of the signal transduction pathways in nearby cells. These multiple intracellular and extracellular pathways of protein modification and signal transduction may constitute the mechanisms by which many of the transient alterations in cellular metabolism occur after exposure to ionizing radiation (Blattner and others 1994).
Some responses observed in particular regimes of exposure to ionizing radiation and given unique names (e.g., adaptive response, bystander effect, genomic instability) may constitute particular manifestations of these general stress responses and signal transduction pathways. These apparently distinct radiation responses have been described mainly in cell biology experiments, and in no case do they have solid biochemical support or mechanistic understanding. In addition to controversy among laboratories, some of the responses described appear to be valid only within a limited dose range and under particular experimental conditions. It is also unclear whether different types of cells, such as epithelial cells, fibroblasts, and lymphoid cells, respond similarly or differently in this regard. Some of the inducible responses appear to be complex in that they depend on participation of intercellular gap junctions in communicating radiation responses to neighboring cells. Work on this subject is in the preliminary, descriptive stage, and there is no understanding of what compounds or factors would be transferred between cells in the gap junction. Therefore, it is difficult to evaluate whether the phenomena are of any general physiologic significance.
In this chapter the committee has provided background information relating to the physical and chemical aspects of radiation and the interaction of radiation with the target molecule DNA. The chapter describes the physics of electrons and beta particles, which are important contributors to direct DNA damage after ionizing radiation exposure, and introduces a special subject—the effect that neutron RBEs have on low-LET radiation risk estimates. Radical formation by ionizing radiation and its contribution to DNA damage are also described. The committee has discussed the contributions of normal oxidative DNA damage relative to radiation-induced DNA damage and described the DNA repair mechanisms that mammalian cells have developed to cope with