Compact form of SV40 viral minichromosome is resistant to nuclease: possible implications for chromatin structure

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Columnar structure of SV40 minichromosome

Like the sequence of the strongest 601 clone nucleosome of Lowary and Widom, the SV40 genome sequence contains tracks of YR dinucleotides separated by small integers of the 10.4n base series (10, 11, 21 and 30 bases). The tracks, however, substantially exceed the nucleosome DNA size and, thus, correspond to more extended structure columnar chromatin. The micrococcal nuclease digests of the SV40...

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Simian virus 40 (SV40) offers an attractive model system for studies on the nucleosomal organization of eukaryotic chromatin [l-3]. SV40 minichromosome is particularly useful for the analysis of the specific nucleosome arrangement on well-defined coding and regulatory sequences of the SV40 genome [4,5]. It has been suggested that the minichromosome contains less nucleosomes than may be expected...

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Non-random cleavage of SV40 DNA in the compact minichromosome and free in solution by micrococcal nuclease.

The distribution of the primary cleavage sites produced by micrococcal nuclease on SV40 DNA in the compact minichromosome was analysed. Minichromosomes purified by improved method were digested in mild conditions in order to minimize possible changes in nucleosomal and supranucleosomal arrangement. The primary cuts were found to be non-randomly and unevenly distributed through the whole SV40 ge...

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Under exhaustive digestion conditions P1 nuclease was found to cleave a subpopulation of intracellular SV40 chromatin only once. The major P1 cleavage site in SV40 DNA was mapped at the origin of DNA replication, and the two minor sites at the SV40 enhancers. The P1-sensitive SV40 chromatin subpopulation was found to have higher superhelical density than the bulk of the intracellular SV40 chrom...

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Micrococcal nuclease analysis of chromatin structure.

This unit describes methodology for using micrococcal nuclease to investigate the presence of nucleosomes at a particular location in chromatin and to map the positions of nucleosomes at various levels of resolution. The approaches are readily adaptable to other probes of chromatin structure that cause DNA cleavage. Results obtained from such chromatin studies provide a structural view of the m...

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ژورنال

عنوان ژورنال: Nucleic Acids Research

سال: 1977

ISSN: 0305-1048,1362-4962

DOI: 10.1093/nar/4.10.3303