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Premature Chromosome Condensation
The present chapter is a review of the types of DNA damage generated under stressful conditions and experimental approaches to the relation of these types of DNA damage to hydroxyurea treatment and caffeine-induced premature chromosome condensation PCC. In this chapter, an attempt is also made to explain the molecular base of DNA damage and to present experimental procedures allowing the illustration of DNA damages at the cell level, especially with the use of histochemical and immunocytochemical methods. It will be experimentally shown, among others, that replication stress mainly leads to the generation of double-strand breaks in DNA DSBs , while the breakage of restrictive interactions of checkpoints during PCC induction results in the accumulation of single-strand breaks SSBs.
DNA can be damaged by the action of endogenous intrinsic or exogenous extrinsic stress factors. The endogenous factors include, among others, errors generated during replication and reactive oxygen species ROS. The exogenous environmental factors are divided into i physical factors, e.
Stress-induced damage includes spontaneous depurination and deamination, oxidation, formation of DNA adducts induced by alkylating agents, formation of cyclobutane dimers, single- and double-strand damage, as well as errors made during replication, repair, reverse transcription and recombination. DNA is also subject to covalent modifications that may affect nitrogen bases and lead to changes in base pairing between DNA strands, or even entirely preventing base pairing.
Genomic instability may also be associated with chromosomal rearrangements which result from changes that occur in the trans position including replication, DNA repair and S phase checkpoint pathways or from changes that act in the cis position, i. Plants, due to their 'settled' lifestyles are exposed to many environmental factors that cause disturbances in the cell cycle.
Each of these burdens leads to the mobilization of defense responses: 1 activation of cell cycle checkpoints and DNA repair factors, 2 inhibition of cell growth, or 3 initiation of the apoptosis pathway Deckert et al. Recognition of double-stranded breaks depends on the MRN complex MreRadNbs1 , necessary for binding chromatin-remodeling factors Schiller et al.