Another potential artifact in the study of nucleosome phasing by chromatin digestion with micrococcal nuclease |
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Authors: | James D McGhee Gary Felsenfeld |
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Institution: | Section on Physical Chemistry Laboratory of Molecular Biology National Institute of Arthritis, Diabetes, and Digestive and Kidney Diseases Bethesda, Maryland 20205 USA |
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Abstract: | We show that, contrary to expectations, restriction enzyme cleavage of chicken erythrocyte nucleosome core particle DNA generates a series of distinct subnucleosome fragments. These fragments do not result from bulk nucleosome phasing in vivo, but arise from micrococcal nuclease cleavages internal to the core particle, at roughly 10-base pair intervals and at AT-rich sequences. Those 145-base pair DNA fragments remaining intact are a biased population in which the guanine content can fluctuate by as much as 10%, with a 10-base pair period. We suggest that these same considerations, when applied to a unique DNA sequence, are the true explanation for several previous claims for nucleosome phasing. |
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