title: Topoisomerase II- and condensin-dependent breakage of MEC1ATR-sensitive fragile sites occurs independently of spindle tension, anaphase, or cytokinesis.
abstract: Fragile sites are loci of recurrent chromosome breakage in the genome. They are found in organisms ranging from bacteria to humans and are implicated in genome instability, evolution, and cancer. In budding yeast, inactivation of Mec1, a homolog of mammalian ATR, leads to chromosome breakage at fragile sites referred to as replication slow zones (RSZs). RSZs are proposed to be homologous to mammalian common fragile sites (CFSs) whose stability is regulated by ATR. Perturbation during S phase, leading to elevated levels of stalled replication forks, is necessary but not sufficient for chromosome breakage at RSZs or CFSs. To address the nature of additional event(s) required for the break formation, we examined involvement of the currently known or implicated mechanisms of endogenous chromosome breakage, including errors in replication fork restart, premature mitotic chromosome condensation, spindle tension, anaphase, and cytokinesis. Results revealed that chromosome breakage at RSZs is independent of the RAD52 epistasis group genes and of TOP3, SGS1, SRS2, MMS4, or MUS81, indicating that homologous recombination and other recombination-related processes associated with replication fork restart are unlikely to be involved. We also found spindle force, anaphase, or cytokinesis to be dispensable. RSZ breakage, however, required genes encoding condensin subunits (YCG1, YSC4) and topoisomerase II (TOP2). We propose that chromosome break formation at RSZs following Mec1 inactivation, a model for mammalian fragile site breakage, is mediated by internal chromosomal stress generated during mitotic chromosome condensation.
authors.affiliation: Division of Stem Cell Biology and Developmental Genetics, National Institute for Medical Research, Medical Research Council, London, United Kingdom.
authors.forename: Nadia
authors.initials: N
authors.lastname: Hashash
authors.forename: Anthony L
authors.initials: AL
authors.lastname: Johnson
authors.forename: Rita S
authors.initials: RS
authors.lastname: Cha
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000705:Anaphase / Q000502:physiology*; D019457:Chromosome Breakage; D043283:Chromosome Fragile Sites; D048749:Cytokinesis / Q000502:physiology*; D004261:DNA Replication; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008938:Mitosis / Q000502:physiology; D046912:Multiprotein Complexes / Q000378:metabolism*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D004718:Saccharomycetales / Q000235:genetics / Q000378:metabolism; D008941:Spindle Apparatus
chemical_list: D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D046912:Multiprotein Complexes; D029701:Saccharomyces cerevisiae Proteins; C108312:condensin complexes; C088937:MEC1 protein, S cerevisiae; D017346:Protein Serine-Threonine Kinases; D000251:Adenosine Triphosphatases; D004250:DNA Topoisomerases, Type II