The chicken lysozyme gene 5' MAR and the Drosophila histone SAR are electroelutable from encapsulated and digested nuclei.
Cultured chicken cells were encapsulated in agarose microbeads, lysed in a near-physiological buffer and resulting encapsulated nuclei were digested with a restriction enzyme and electroeluted. After removal of approximately 97% of the chromatin, the nuclear lamina, residual nucleoli and an internal nuclear network remained. The majority of nascent RNA was also recovered in digested and electroeluted nuclei. Surprisingly, however, the chicken lysozyme gene 5' MAR was quantitatively electroeluted from digested nuclei of expressing and non-expressing cells, as well as the promoter region and the coding sequence. When encapsulated nuclei were digested partially, the proportion of elutable 5' MAR chromatin was comparable to that of elutable bulk chromatin. Furthermore, after digestion of encapsulated nuclei from Drosophila Kc cells, the histone SAR was electroeluted to the same extent as bulk chromatin. We conclude that the lysozyme gene 5' MAR and the histone SAR are not permanently attached to a nuclear matrix or scaffold.
Hempel
K
K
Institut für Physiologische Chemie, Universitäts-Krankenhaus Eppendorf, Hamburg, FR Germany.
Strätling
W H
WH
D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry / Q000648:ultrastructure; D002645:Chickens; D002843:Chromatin / Q000302:isolation & purification; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D004330:Drosophila; D006657:Histones / Q000302:isolation & purification*; D008854:Microscopy, Electron; D008863:Microspheres; D009113:Muramidase / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012685:Sepharose
D002843:Chromatin; D006657:Histones; D004247:DNA; D012685:Sepharose; D009113:Muramidase