title: Chromatin dynamics in living cells: identification of oscillatory motion.
abstract: Genomic DNA in mammalian cells is organized into ~1 Mbp chromatin domains (ChrD) which represent the basic structural units for DNA compaction, replication, and transcription. Remarkably, ChrD are highly dynamic and undergo both translational movement and configurational changes. In this study, we introduce an automated motion tracking analysis to measure, both in 2D and 3D, the linear displacement of early, mid and late S-phase replicated ChrD over short time periods (<1 sec). We conclude that previously identified large-scale transitions in the spatial position and configuration of chromatin, originate from asymmetric oscillations of the ChrD detectable in fractions of a second. The rapid oscillatory motion correlates with the replication timing of the ChrD with early S replicated ChrD showing the highest levels of motion and late S-phase chromatin the lowest. Virtually identical levels of oscillatory motion were detected when ChrD were measured during active DNA replication or during inhibition of transcription with DRB or α-amanitin. While this motion is energy independent, the oscillations of early S and mid S, but not late S replicated chromatin, are reduced by cell permeabilization. This suggests involvement of soluble factors in the regulation of chromatin dynamics. The DNA intercalating agent actinomycin D also significantly inhibits early S-labeled chromatin oscillation. We propose that rapid asymmetric oscillations of <1 sec are the basis for translational movements and configurational changes in ChrD previously detected over time spans of minutes-hours, and are the result of both the stochastic collisions of macromolecules and specific molecular interactions.
authors.affiliation: Department of Biological Sciences, University at Buffalo, The State University of New York, Buffalo, New York 14260, USA.
authors.forename: Artem
authors.initials: A
authors.lastname: Pliss
authors.forename: Kishore S
authors.initials: KS
authors.lastname: Malyavantham
authors.forename: Sambit
authors.initials: S
authors.lastname: Bhattacharya
authors.forename: Ronald
authors.initials: R
authors.lastname: Berezney
mesh_terms: D002463:Cell Membrane Permeability; D002843:Chromatin / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D004261:DNA Replication; D005456:Fluorescent Dyes; D006367:HeLa Cells; D006801:Humans; D021621:Imaging, Three-Dimensional; D046911:Macromolecular Substances; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009068:Movement / Q000502:physiology; D016196:S Phase; D013269:Stochastic Processes
chemical_list: D002843:Chromatin; D005456:Fluorescent Dyes; D046911:Macromolecular Substances; D004247:DNA