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The connection between chromatin motion on the 100 nm length scale and core histone dynamics in live XTC-2 cells and isolated nuclei.
Davis SK
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Bardeen CJ
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The diffusive motion of DNA-containing chromatin in live cells and isolated nuclei is investigated using a two-photon standing wave fluorescence photobleaching experiment with 100 nm spatial resolution. The chromatin is labeled using the minor groove binding dye Hoechst 33342. In live cells, the mean diffusion rate is 5 x 10(-4) micro m2/s, with considerable cell-to-cell variation. This diffusion is highly constrained and cannot be observed in a standard, single beam fluorescence recovery after photobleaching experiment. To determine the chemical origin of the diffusion, we study motion in isolated nuclei and vary the strength of the histone-DNA interactions by changing the ionic strength and using chemical and photocross-linking experiments. At higher NaCl concentrations, we see increased chromatin diffusion as the histone-DNA interaction is weakened due to ionic screening, whereas photocross-linking the core histones to the DNA results in a complete absence of diffusive motion. These trends are consistent with the 100 nm scale motion being correlated with the interactions of histone proteins with the DNA. If chromatin diffusion is connected to the nucleosomal dynamics on much smaller length scales, this may provide a way to assay biochemical activity in vivo based on larger scale macromolecular dynamics observed via fluorescence microscopy.
Abney,
Chromatin dynamics in interphase nuclei and its implications for nuclear structure.
1997, Pubmed
Abney,
Chromatin dynamics in interphase nuclei and its implications for nuclear structure.
1997,
Pubmed
Bailey,
Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation.
1993,
Pubmed
Becker,
Nucleosome sliding: facts and fiction.
2002,
Pubmed
Bornfleth,
Quantitative motion analysis of subchromosomal foci in living cells using four-dimensional microscopy.
1999,
Pubmed
Caserta,
Aspects of nucleosomal positional flexibility and fluidity.
2002,
Pubmed
Chubb,
Chromatin motion is constrained by association with nuclear compartments in human cells.
2002,
Pubmed
Clark,
Electrostatic mechanism of chromatin folding.
1990,
Pubmed
Cremer,
Chromosome territories, nuclear architecture and gene regulation in mammalian cells.
2001,
Pubmed
Davis,
Cross-linking of histone proteins to DNA by UV illumination of chromatin stained with Hoechst 33342.
2003,
Pubmed
,
Xenbase
Davoust,
Fringe pattern photobleaching, a new method for the measurement of transport coefficients of biological macromolecules.
1982,
Pubmed
Durand,
Cytotoxicity, Mutagenicity and DNA damage by Hoechst 33342.
1982,
Pubmed
Gerchman,
Chromatin higher-order structure studied by neutron scattering and scanning transmission electron microscopy.
1987,
Pubmed
Gunawardena,
Direct evidence for interphase chromosome movement during the mid-blastula transition in Drosophila.
2000,
Pubmed
Guschin,
ATP-Dependent histone octamer mobilization and histone deacetylation mediated by the Mi-2 chromatin remodeling complex.
2000,
Pubmed
,
Xenbase
Guschin,
Multiple ISWI ATPase complexes from xenopus laevis. Functional conservation of an ACF/CHRAC homolog.
2000,
Pubmed
,
Xenbase
Gustafsson,
I5M: 3D widefield light microscopy with better than 100 nm axial resolution.
1999,
Pubmed
Hamiche,
ATP-dependent histone octamer sliding mediated by the chromatin remodeling complex NURF.
1999,
Pubmed
Hansen,
Conformational dynamics of the chromatin fiber in solution: determinants, mechanisms, and functions.
2002,
Pubmed
Hell,
Far-field fluorescence microscopy with three-dimensional resolution in the 100-nm range.
1997,
Pubmed
Heun,
Chromosome dynamics in the yeast interphase nucleus.
2001,
Pubmed
Jackson,
Formaldehyde cross-linking for studying nucleosomal dynamics.
1999,
Pubmed
Kanda,
Histone-GFP fusion protein enables sensitive analysis of chromosome dynamics in living mammalian cells.
1998,
Pubmed
Kimura,
Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B.
2001,
Pubmed
Lamond,
Structure and function in the nucleus.
1998,
Pubmed
Lever,
Rapid exchange of histone H1.1 on chromatin in living human cells.
2000,
Pubmed
Loontiens,
Binding characteristics of Hoechst 33258 with calf thymus DNA, poly[d(A-T)], and d(CCGGAATTCCGG): multiple stoichiometries and determination of tight binding with a wide spectrum of site affinities.
1990,
Pubmed
Luger,
Structure and dynamic behavior of nucleosomes.
2003,
Pubmed
,
Xenbase
Marshall,
Interphase chromosomes undergo constrained diffusional motion in living cells.
1997,
Pubmed
Marshall,
Order and disorder in the nucleus.
2002,
Pubmed
Meersseman,
Mobile nucleosomes--a general behavior.
1992,
Pubmed
Misteli,
Dynamic binding of histone H1 to chromatin in living cells.
2000,
Pubmed
Phair,
High mobility of proteins in the mammalian cell nucleus.
2000,
Pubmed
Sadoni,
Large-scale chromatin fibers of living cells display a discontinuous functional organization.
2001,
Pubmed
Schlax,
Inhibition of transcription initiation by lac repressor.
1995,
Pubmed
Selvin,
A polarized photobleaching study of chromatin reorientation in intact nuclei.
1990,
Pubmed
Siino,
Photobleaching of GFP-labeled H2AX in chromatin: H2AX has low diffusional mobility in the nucleus.
2002,
Pubmed
Smith,
Determination of molecular motion in membranes using periodic pattern photobleaching.
1978,
Pubmed
Smith,
Involvement of DNA topoisomerase II in the selective resistance of a mammalian cell mutant to DNA minor groove ligands: ligand-induced DNA-protein crosslinking and responses to topoisomerase poisons.
1990,
Pubmed
Spadafora,
Rearrangement of chromatin structure induced by increasing ionic strength and temperature.
1979,
Pubmed
Tong,
Chromatin deacetylation by an ATP-dependent nucleosome remodelling complex.
1998,
Pubmed
Tsukamoto,
Visualization of gene activity in living cells.
2000,
Pubmed
Tumbar,
Large-scale chromatin unfolding and remodeling induced by VP16 acidic activation domain.
1999,
Pubmed
Varga-Weisz,
Energy-dependent chromatin accessibility and nucleosome mobility in a cell-free system.
1995,
Pubmed
Vazquez,
Multiple regimes of constrained chromosome motion are regulated in the interphase Drosophila nucleus.
2001,
Pubmed
Vermaak,
Chromatin and chromosomal controls in development.
1998,
Pubmed
Weischet,
On the de novo formation of compact oligonucleosomes at high ionic strength. Evidence for nucleosomal sliding in high salt.
1979,
Pubmed
Widom,
Structure, dynamics, and function of chromatin in vitro.
1998,
Pubmed
Wolffe,
Nuclear visions: functional flexibility from structural instability.
2001,
Pubmed