Chromatin: Proteine im Zellkern

Cystals have been obtained on nucleosome cores and analysed by X-ray diffraction and electron microscopy. The core is a flat particle of dimensions about 110 X 110 X 57 A, somewhat wedge shaped, and strongly divided into two 'layers', consistent with the DNA being wound into about 1 3/4 turns of a flat superhelix of a pitch about 28 A. The organisation of the DNA can be correlated with the results to enzyme digestion studies. A change in the screw of the DNA double helix on nucleosome formation can be deduced.

Biochemical evidence is presented which confirms that the DNA repeat length in micrococcal nuclease (spleen endonuclease, nucleate 3'-oligonucleotidohydrolase, EC 3-1-4-7) digests of Chinese hamster ovary chromatin is shorter than that of rat liver chromatin [J.L. Compton, R. Hancock, P. Oudet, and P. Chambon (1976) Eur. J. Biochem., in press]. A survey of available cells has shown that the DNA repeat length of the chromatin of higher eukaryotes varies widely. A value of 196 base pairs was found for cells of all mature tissues, regardless of the source of the tissue, whereas smaller values were found for cells of actively dividing tissues and larger values were found for a genetically inactive cell. Although the DNA repeat length of the chromatin of cells in culture was usually shorter than 196 base pairs, there was no general correlation between the size of the chromatin DNA repeat length and the rate of cell division or the functional state of the cell in culture. Examination of extensive micrococcal nuclease digests suggests that the chromatin subunits of all of the higher eukaryotic cells we have studied contain a core with approximately 140 base pairs of DNA.

Two major nonhistone polypeptides from rat liver chromatin have been identified as myosin and actin. Preliminary observations indicate that three other chromatin polypeptides of molecular weights 50,000, 34,000, and 32,000 are tubulin and heavy and light tropomyosin, respectively. A sixth component of molecular weight 65,000 which has been purified and electrophoreses as a single band on sodium dodecyl sulfate-polyacrylamide gels may be composed in part of protease-digested myosin. These six polypeptides together account for as much as 38% of the nonhistone protein mass of chromatin in this tissue.