[Comparative analysis of subdomain fragments of chromatin].
Nuclei isolated from Djungarian hamster fibroblasts transformed by SV40 were treated with restriction endonuclease Bsp RI, fixed on Celite columns and underwent successive gradients of dissociating agents, such as NaCl, LiCl-urea, and temperature. This procedure leads to fractionation of DNA fragments in accordance with the tightness of DNA-protein bonds in situ. The fractions obtained were analysed by agarose gel electrophoresis and dot-hybridization technique with the use of various DNA probes. The results received are as follows: a) a DNA fragment size is not a factor determining the chromatographic position, the latter is probably stipulated by DNA-protein interactions; b) an analysis of cells synchronized at the G1/S border shows that the distribution of specific DNA sequences, such as actin, histone, hsp 70, and c-Ha-ras genes as well as reiterated DNA sequences, does not coincide with that of total genomic DNA; the nuclear matrix-attached fragments of those sequences are enriched to various extents. By nick-translation labeling in situ, DNase I-sensitive and hypersensitive regions were tentatively identified among subdomain chromatin fragments.
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D000818:Animals; D002461:Cell Line, Transformed; D002843:Chromatin / Q000378:metabolism*; D002853:Chromatography, Liquid; D006224:Cricetinae; D004247:DNA / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D005347:Fibroblasts / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D011506:Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship
D002843:Chromatin; D011506:Proteins; D004247:DNA
Sravnitel'nyĭ analiz subdomennykh fragmentov khromatina.