Large-scale chromatin structural domains within mitotic and interphase chromosomes in vivo and in vitro (original) (raw)
Abstract
Higher-order chromatin structural domains approximately 130 nm in width are observed as prominent components of both Drosophila melanogaster and human mitotic chromosomes using buffer conditions which preserve chromosome morphology as determined by light microscopic comparison with chromosomes within living cells. Spatially discrete chromatin structural domains of similar size also exist as prominent components within interphase nuclei prepared under equivalent conditions. Examination of chromosomes during the anaphase-telophase transition suggests that chromosomes decondense largely through the progressive straightening or uncoiling of these large-scale chromatin domains. A quantitative analysis of the size distribution of these higher-order domains in telophase nuclei indicated a mean width of 126±36 nm. Three-dimensional views using stereopairs of chromosomes and interphase nuclei from 0.5 μm thick sections suggest that these large-scale chromatin domains consist of 30 nm fibers packed by tight folding into larger, linear, fiber-like elements. Reduction in vitro of either polyamine or divalent cation concentrations within two different buffer systems results in a loss of these large-scale domains, with no higher-order chromatin organization evident above the 20–30 nm fiber. Under these conditions the DNA distribution within mitotic chromosomes and interphase nuclei appears significantly diffuse relative to the appearance by light microscopy within living cells, or, by electron microscopy, within cells fixed directly without permeabilization in buffer. These results suggest that these large-scale chromatin structural domains are fundamental elements of chromosome architecture in vivo.
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- Andrew S. Belmont
Present address: Department of Cell and Structural Biology, University of Illinois, 61801, Urbana, Urbana, IL, USA
Authors and Affiliations
- Department of Biochemistry and Biophysics and the Howard Hughes Medical Institute, University of California, 94143-0448, San Francisco, San Francisco, CA, USA
Andrew S. Belmont, Michael B. Braunfeld, John W. Sedat & David A. Agard
Authors
- Andrew S. Belmont
You can also search for this author inPubMed Google Scholar - Michael B. Braunfeld
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Belmont, A.S., Braunfeld, M.B., Sedat, J.W. et al. Large-scale chromatin structural domains within mitotic and interphase chromosomes in vivo and in vitro.Chromosoma 98, 129–143 (1989). https://doi.org/10.1007/BF00291049
- Received: 01 December 1988
- Revised: 02 March 1989
- Issue Date: August 1989
- DOI: https://doi.org/10.1007/BF00291049