@article{ScheerSommervilleMueller1980, author = {Scheer, Ulrich and Sommerville, John and M{\"u}ller, Ulrike}, title = {DNA is assembled into globular supranucleosomal chromatin structures by nuclear contents of amphibian oocytes}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-39671}, year = {1980}, abstract = {The assembly of DNA into nucleosomal and supranucleosomal chromatin structures has been studied (i) by injection of circular DNA molecules (plasmids) into nuclei of Pleurodeles waltlii oocytes; and (ii) by in vitro incubation of plasmid molecules with the supernatant fraction from oocyte nuclei of Pleurodeles and Xenopus laevis, followed by purification of nucleoprotein structures formed with sucrose gradient centrifugation. [n both types of experiments , spread preparations of the newly assembled and transcriptionally inactive chromatin , examined by electron microscopy , show dense globular higher order (supranucleosomal) packing forms. Under partially relaxing (low salt) preparation conditions granular chromatin subunits of about 30 nm diameter can be seen either as widely spaced particles or in closely packed aggregates. The transcriptionally inactive endogenous chromatin of chromomeres of lampbrush chromosomes is arranged in similar higher order chromatin units. A correlation is found between the sizes of the DN A molecule probes used and the numbers of nucleosomes and higher order globules in the assembled chromatin structures. After prolonged dispersion in low salt buffers , these globular chromatin units unfold into chains of7-12 nucleosomes. The results support the concept that chromatin is arranged , under physiological ion concentrations as they are present in the nucleus , in supranucleosomal units of globular morphology.}, language = {en} } @article{ZentgrafTrendelenburgSpringetal.1979, author = {Zentgraf, Hanswalter and Trendelenburg, Michael F. and Spring, Herbert and Scheer, Ulrich and Franke, Werner W. and M{\"u}ller, Ulrike and Drury, Kenneth C. and Rungger, Duri}, title = {Mitochondrial DNA arranged into chromatin-like structures after injection into amphibian oocyte nuclei}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-33174}, year = {1979}, abstract = {Purified mitochondrial DNA (mitDNA) from ovaries ofXenopus lae vis was injected into the nuclei (germinal vesicles) of large viteUogenic oocytes of the same organism and examined by electron microscopy ofthe spread nuclear contents. Normally located nuclei of untreated oocytes as weil as peripherally translocated nuclei of centrifuged oocytes were used. In addition, oocyte nuclei isolated and incubated under liquid paraffin oil were injected with DNA. The integrity oftranscriptional structures of endogenous chromosomal (Iampbrush chromosomes) and extrachromosomal (nucleoli) genes of the injected nuclei was demonstrated. Microinjected mitDN A was identified as circles of chromatin exhibiting polynucleosome-like organization and a me an contour length of 2.6 J.Lm, corresponding to a compaction ratio of the mitDN A of about 2 : I. This DNA packing ratio is similar to that observed after preparation of various kinds of native chromatin in low salt buffers. The chromatin circles formed from injected mitDNA only very rarely exhibited lateral fibrils suggestive of transcriptional activity. These results suggest that purified mitDNA can be transformed to normally structured chromatin when exposed to oocyte nuclear contents but is rarely , if at all , transcribed in this form and in this environment.}, language = {en} }