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DNA Dynamics during Early Double-Strand Break Processing Revealed by Non-Intrusive Imaging of Living Cells


When chromosomes break, cells must repair them to avoid becoming abnormal, cancerous or dead. The most accurate repair mechanism is based on homologous recombination (HR), in which single strands generated next to the break seek an intact replica which is copied into the broken site. Changes in chromosome dynamics during the early steps that create the single strands have not been analysed owing to lack of tools allowing analysis of this process in individual living cells. We have developed a method for directly observing the resection process that prepares DNA double-strand breaks for HR. This allows us for the first time to identify just those cells where breaks are being repaired, and so to analyze the repair mechanism with a precision not attainable using current visualization systems. We have observed that the broken DNA is prepared for restoration much faster than previously thought, and that DNA movement first slows dramatically, prior to the more pronounced movement previously seen to accompany later stages of repair.


Vyšlo v časopise: DNA Dynamics during Early Double-Strand Break Processing Revealed by Non-Intrusive Imaging of Living Cells. PLoS Genet 10(3): e32767. doi:10.1371/journal.pgen.1004187
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1004187

Souhrn

When chromosomes break, cells must repair them to avoid becoming abnormal, cancerous or dead. The most accurate repair mechanism is based on homologous recombination (HR), in which single strands generated next to the break seek an intact replica which is copied into the broken site. Changes in chromosome dynamics during the early steps that create the single strands have not been analysed owing to lack of tools allowing analysis of this process in individual living cells. We have developed a method for directly observing the resection process that prepares DNA double-strand breaks for HR. This allows us for the first time to identify just those cells where breaks are being repaired, and so to analyze the repair mechanism with a precision not attainable using current visualization systems. We have observed that the broken DNA is prepared for restoration much faster than previously thought, and that DNA movement first slows dramatically, prior to the more pronounced movement previously seen to accompany later stages of repair.


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