Abstract
The higher-order structure of chromatin isolated from sea urchin sperm, which has a long nucleosomal DNA repeat length (approximately 240 bp), has been studied by electron microscopy and X-ray diffraction. Electron micrographs show that this chromatin forms 300 A filaments which are indistinguishable from those of chicken erythrocytes (approximately 212 bp repeat); X-ray diffraction patterns from partially oriented samples show that the edge-to-edge packing of nucleosomes in the direction of the 300 A filament axis, and the radial disposition of nucleosomes around it, are both similar to those of the chicken erythrocyte 300 A filament, which is described by the solenoid model. The invariance of the structure with increased linker DNA length is inconsistent with many other models proposed for the 300 A filament and, furthermore, means that the linker DNA must be bent. The low-angle X-ray scattering in the 300-400 A region both in vitro and in vivo differs from that of chicken erythrocyte chromatin. The nature of the difference suggests that 300 A filaments in sea urchin sperm in vivo are packed so tightly together that electron-density contrast between individual filaments is lost; this is consistent with electron micrographs of the chromatin in vitro.
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Selected References
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- Allan J., Rau D. C., Harborne N., Gould H. Higher order structure in a short repeat length chromatin. J Cell Biol. 1984 Apr;98(4):1320–1327. doi: 10.1083/jcb.98.4.1320. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ausio J., Borochov N., Seger D., Eisenberg H. Interaction of chromatin with NaCl and MgCl2. Solubility and binding studies, transition to and characterization of the higher-order structure. J Mol Biol. 1984 Aug 15;177(3):373–398. doi: 10.1016/0022-2836(84)90291-2. [DOI] [PubMed] [Google Scholar]
- Bates D. L., Butler P. J., Pearson E. C., Thomas J. O. Stability of the higher-order structure of chicken-erythrocyte chromatin in solution. Eur J Biochem. 1981 Oct;119(3):469–476. doi: 10.1111/j.1432-1033.1981.tb05631.x. [DOI] [PubMed] [Google Scholar]
- Butler P. J. A defined structure of the 30 nm chromatin fibre which accommodates different nucleosomal repeat lengths. EMBO J. 1984 Nov;3(11):2599–2604. doi: 10.1002/j.1460-2075.1984.tb02180.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Butler P. J. The folding of chromatin. CRC Crit Rev Biochem. 1983;15(1):57–91. doi: 10.3109/10409238309102801. [DOI] [PubMed] [Google Scholar]
- Butler P. J., Thomas J. O. Changes in chromatin folding in solution. J Mol Biol. 1980 Jul 15;140(4):505–529. doi: 10.1016/0022-2836(80)90268-5. [DOI] [PubMed] [Google Scholar]
- Finch J. T., Klug A. Solenoidal model for superstructure in chromatin. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1897–1901. doi: 10.1073/pnas.73.6.1897. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hewish D. R., Burgoyne L. A. Chromatin sub-structure. The digestion of chromatin DNA at regularly spaced sites by a nuclear deoxyribonuclease. Biochem Biophys Res Commun. 1973 May 15;52(2):504–510. doi: 10.1016/0006-291x(73)90740-7. [DOI] [PubMed] [Google Scholar]
- Kornberg R. D. Structure of chromatin. Annu Rev Biochem. 1977;46:931–954. doi: 10.1146/annurev.bi.46.070177.004435. [DOI] [PubMed] [Google Scholar]
- Langmore J. P., Paulson J. R. Low angle x-ray diffraction studies of chromatin structure in vivo and in isolated nuclei and metaphase chromosomes. J Cell Biol. 1983 Apr;96(4):1120–1131. doi: 10.1083/jcb.96.4.1120. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Langmore J. P., Schutt C. The higher order structure of chicken erythrocyte chromosomes in vivo. Nature. 1980 Dec 11;288(5791):620–622. doi: 10.1038/288620a0. [DOI] [PubMed] [Google Scholar]
- Marsden M. P., Laemmli U. K. Metaphase chromosome structure: evidence for a radial loop model. Cell. 1979 Aug;17(4):849–858. doi: 10.1016/0092-8674(79)90325-8. [DOI] [PubMed] [Google Scholar]
- McGhee J. D., Nickol J. M., Felsenfeld G., Rau D. C. Higher order structure of chromatin: orientation of nucleosomes within the 30 nm chromatin solenoid is independent of species and spacer length. Cell. 1983 Jul;33(3):831–841. doi: 10.1016/0092-8674(83)90025-9. [DOI] [PubMed] [Google Scholar]
- McGhee J. D., Rau D. C., Charney E., Felsenfeld G. Orientation of the nucleosome within the higher order structure of chromatin. Cell. 1980 Nov;22(1 Pt 1):87–96. doi: 10.1016/0092-8674(80)90157-9. [DOI] [PubMed] [Google Scholar]
- Mitra S., Sen D., Crothers D. M. Orientation of nucleosomes and linker DNA in calf thymus chromatin determined by photochemical dichroism. Nature. 1984 Mar 15;308(5956):247–250. doi: 10.1038/308247a0. [DOI] [PubMed] [Google Scholar]
- Morris N. R. A comparison of the structure of chicken erythrocyte and chicken liver chromatin. Cell. 1976 Dec;9(4 Pt 1):627–632. doi: 10.1016/0092-8674(76)90045-3. [DOI] [PubMed] [Google Scholar]
- Noll M., Thomas J. O., Kornberg R. D. Preparation of native chromatin and damage caused by shearing. Science. 1975 Mar 28;187(4182):1203–1206. doi: 10.1126/science.187.4182.1203. [DOI] [PubMed] [Google Scholar]
- Pearson E. C., Butler P. J., Thomas J. O. Higher-order structure of nucleosome oligomers from short-repeat chromatin. EMBO J. 1983;2(8):1367–1372. doi: 10.1002/j.1460-2075.1983.tb01593.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spadafora C., Bellard M., Compton J. L., Chambon P. The DNA repeat lengths in chromatins from sea urchin sperm and gastrule cells are markedly different. FEBS Lett. 1976 Oct 15;69(1):281–285. doi: 10.1016/0014-5793(76)80704-1. [DOI] [PubMed] [Google Scholar]
- Strätling W. H., Klingholz R. Supranucleosomal structure of chromatin: digestion by calcium/magnesium endonuclease proceeds via a discrete size class of particles with elevated stability. Biochemistry. 1981 Mar 3;20(5):1386–1392. doi: 10.1021/bi00508a054. [DOI] [PubMed] [Google Scholar]
- Suau P., Bradbury E. M., Baldwin J. P. Higher-order structures of chromatin in solution. Eur J Biochem. 1979 Jul;97(2):593–602. doi: 10.1111/j.1432-1033.1979.tb13148.x. [DOI] [PubMed] [Google Scholar]
- Thoma F., Koller T., Klug A. Involvement of histone H1 in the organization of the nucleosome and of the salt-dependent superstructures of chromatin. J Cell Biol. 1979 Nov;83(2 Pt 1):403–427. doi: 10.1083/jcb.83.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas J. O., Butler P. J. Size-dependence of a stable higher-order structure of chromatin. J Mol Biol. 1980 Nov 25;144(1):89–93. doi: 10.1016/0022-2836(80)90215-6. [DOI] [PubMed] [Google Scholar]
- Thomas J. O., Kornberg R. D. The study of histone--histone associations by chemical cross-linking. Methods Cell Biol. 1978;18:429–440. [PubMed] [Google Scholar]
- Thomas J. O., Rees C., Pearson E. C. Histone H5 promotes the association of condensed chromatin fragments to give pseudo-higher-order structures. Eur J Biochem. 1985 Feb 15;147(1):143–151. doi: 10.1111/j.1432-1033.1985.tb08730.x. [DOI] [PubMed] [Google Scholar]
- Walmsley M. E., Davies H. G. Ultrastructural and biochemical observations on interphase nuclei isolated from chicken erythrocytes. J Cell Sci. 1975 Jan;17(1):113–139. doi: 10.1242/jcs.17.1.113. [DOI] [PubMed] [Google Scholar]
- Woodcock C. L., Frado L. L., Rattner J. B. The higher-order structure of chromatin: evidence for a helical ribbon arrangement. J Cell Biol. 1984 Jul;99(1 Pt 1):42–52. doi: 10.1083/jcb.99.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Worcel A., Strogatz S., Riley D. Structure of chromatin and the linking number of DNA. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1461–1465. doi: 10.1073/pnas.78.3.1461. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yabuki H., Dattagupta N., Crothers D. M. Orientation of nucleosomes in the thirty-nanometer chromatin fiber. Biochemistry. 1982 Sep 28;21(20):5015–5020. doi: 10.1021/bi00263a027. [DOI] [PubMed] [Google Scholar]
- Zentgraf H., Franke W. W. Differences of supranucleosomal organization in different kinds of chromatin: cell type-specific globular subunits containing different numbers of nucleosomes. J Cell Biol. 1984 Jul;99(1 Pt 1):272–286. doi: 10.1083/jcb.99.1.272. [DOI] [PMC free article] [PubMed] [Google Scholar]