The organization of supercoiled DNA from human chromosomes. 1979

A M Mullinger, and R T Johnson

Nucleoids obtained from metaphase-arrested HeLa cells by gentle lysis in non-ionic detergent and 2 M salt were prepared for electron microscopy by Kleinschmidt spreading techniques. From biophysical studies nucleoids have been shown to contain high molecular weight DNA which is supercoiled and topologically constrained. The contents of nucleoids disperse and collapse into 2 dimensions on the water surface as discrete particles (spreads), which are provisionally equated with metaphase chromosomes. In some cases separate spreads are linked. The structure of the spreads is complex. A preliminary description is presented, although we believe that some of the structures reported may be products of preparation and spreading rather than significant features of the organization of intact nucleoids. There is considerable variation in the appearance of different spreads which may be related to the degree of unfolding, spreading or damage of the different preparations. Each spread consists of one or more core areas surrounded by a network of fibres. Cores are composed, at least in part, of compressed fibres and superficially appear to constrain fibres of the network. In the network the inter-twining of pairs of fibres and the occurrence of flat fibre spirals (disks) are interpreted as evidence of DNA supercoiling, but other fibres of similar thickness are not visibly supercoiled. High-order structures including groupings of disks are observed: these may be the structural correlates of the folded domains of supercoiled DNA established biophysically. The relative proportion of disks and other network structures varies between spreads.

UI MeSH Term Description Entries
D008854 Microscopy, Electron Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen. Electron Microscopy
D002877 Chromosomes, Human Very long DNA molecules and associated proteins, HISTONES, and non-histone chromosomal proteins (CHROMOSOMAL PROTEINS, NON-HISTONE). Normally 46 chromosomes, including two sex chromosomes are found in the nucleus of human cells. They carry the hereditary information of the individual. Chromosome, Human,Human Chromosome,Human Chromosomes
D004278 DNA, Superhelical Circular duplex DNA isolated from viruses, bacteria and mitochondria in supercoiled or supertwisted form. This superhelical DNA is endowed with free energy. During transcription, the magnitude of RNA initiation is proportional to the DNA superhelicity. DNA, Supercoiled,DNA, Supertwisted,Supercoiled DNA,Superhelical DNA,Supertwisted DNA
D006367 HeLa Cells The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for, among other things, VIRUS CULTIVATION and PRECLINICAL DRUG EVALUATION assays. Cell, HeLa,Cells, HeLa,HeLa Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man

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