RRC ID 19619
Author Petrova B, Dehler S, Kruitwagen T, Hériché JK, Miura K, Haering CH.
Title Quantitative analysis of chromosome condensation in fission yeast.
Journal Mol Cell Biol
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.
Volume 33(5)
Pages 984-98
Published 2013-3-1
DOI 10.1128/MCB.01400-12
PII MCB.01400-12
PMID 23263988
PMC PMC3623070
MeSH Acetylation Adenosine Triphosphatases / metabolism Aurora Kinases Chromosomes, Fungal / metabolism* Chromosomes, Fungal / ultrastructure* DNA Topoisomerases, Type II / metabolism DNA-Binding Proteins / metabolism Histones / metabolism Image Processing, Computer-Assisted Meiosis Microscopy* / methods Mitosis Multiprotein Complexes / metabolism Protein Serine-Threonine Kinases / metabolism Schizosaccharomyces / cytology* Schizosaccharomyces / metabolism Schizosaccharomyces pombe Proteins / metabolism
IF 3.611
Times Cited 32