title: Quantitative analysis of chromosome condensation in fission yeast.
abstract: Chromosomes undergo extensive conformational rearrangements in preparation for their segregation during cell divisions. Insights into the molecular mechanisms behind this still poorly understood condensation process require the development of new approaches to quantitatively assess chromosome formation in vivo. In this study, we present a live-cell microscopy-based chromosome condensation assay in the fission yeast Schizosaccharomyces pombe. By automatically tracking the three-dimensional distance changes between fluorescently marked chromosome loci at high temporal and spatial resolution, we analyze chromosome condensation during mitosis and meiosis and deduct defined parameters to describe condensation dynamics. We demonstrate that this method can determine the contributions of condensin, topoisomerase II, and Aurora kinase to mitotic chromosome condensation. We furthermore show that the assay can identify proteins required for mitotic chromosome formation de novo by isolating mutants in condensin, DNA polymerase ε, and F-box DNA helicase I that are specifically defective in pro-/metaphase condensation. Thus, the chromosome condensation assay provides a direct and sensitive system for the discovery and characterization of components of the chromosome condensation machinery in a genetically tractable eukaryote.
authors.affiliation: Cell Biology & Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Boryana
authors.initials: B
authors.lastname: Petrova
authors.forename: Sascha
authors.initials: S
authors.lastname: Dehler
authors.forename: Tom
authors.initials: T
authors.lastname: Kruitwagen
authors.forename: Jean-Karim
authors.initials: JK
authors.lastname: Hériché
authors.forename: Kota
authors.initials: K
authors.lastname: Miura
authors.forename: Christian H
authors.initials: CH
authors.lastname: Haering
mesh_terms: D000107:Acetylation; D000251:Adenosine Triphosphatases / Q000378:metabolism; D064127:Aurora Kinases; D015825:Chromosomes, Fungal / Q000378:metabolism* / Q000648:ultrastructure*; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D008540:Meiosis; D008853:Microscopy / Q000379:methods; D008938:Mitosis; D046912:Multiprotein Complexes / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D012568:Schizosaccharomyces / Q000166:cytology* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D006657:Histones; D046912:Multiprotein Complexes; D029702:Schizosaccharomyces pombe Proteins; C108312:condensin complexes; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; D000251:Adenosine Triphosphatases; D004250:DNA Topoisomerases, Type II