Chromatin structure : possible regulatory role in gene expression Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/44558h24p

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  • Chromatin DNA sequences intimately associated with the "core" of the basic, repeating chromatin subunit are shown to be relatively resistant to digestion by micrococcal nuclease. Chromatin DNA sequences lying between adjacent chromatin subunits, as well as DNA sequences in regions of chromatin which are free of chromatin subunits, are subject to micrococcal nuclease digestion. Thus, the susceptibility of a given sequence of chromatin DNA to nuclease digestion is dependent upon the structural features of the chromatin in which it is located. Nuclease digestion of chromatin is used, together with nucleic acid hybridization, to determine the nucleasesusceptibility of DNA sequences complementary to those of the RNA tumor viruses integrated into the genomes of normal and virus-infected cells. The susceptibility of a given DNA sequence to nuclease digestion is taken to reflect the structure of the region of chromatin in which it is located. By selecting cells in which the virus-specific DNA sequences were present in different states of transcriptional activity, it was possible to determine if differences exist between transcriptionally- active and inactive regions of chromatin. Evidence is presented to demonstrate that the transcriptionally inactive, endogenous, viral DNA sequences in normal chicken erythrocytes are equally susceptible to micrococcal nuclease digestion as are the bulk of the transcriptionally inactive cellular DNA sequences. At the same time, the transcriptionally active viral DNA sequences in leukemic myeloblasts are shown to be more susceptible to micrococcal nuclease digestion than the bulk of the transcriptionally inactive DNA sequences in this cell. The greater nuclease-susceptibility of the viral DNA sequences in leukemic myeloblasts is thought to reflect the different structural organization of that region of chromatin in which they are located. The results reported here demonstrate that structural differences exist between transcriptionally active and inactive regions of chromatin. As a result, support is given to the concept that the structural organization of eucaryotic chromatin may be involved in the regulation of selective gene expression.
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