Nuclease digestion studies of chromatin structure
Micrococcal nuclease, which preferentially cleaves linker DNA in chromatin, was immobilized by covalent attachment to CNBr-activated agarose beads and used to study the accessibility of linker DNA in chromatin fibers prepared from chicken erythrocyte nuclei. This immobilized nuclease was able to cleave chromatin fibers into the typical pattern of fragments corresponding to multiples of mononucleosomes. Cleavage from only the ends of the fibers was ruled out by examining the products of cleavage of fibers end-labelled with /sup 35/P. Comparison of the rate of digestion by immobilized and soluble micrococcal nuclease indicated that the fiber structure does not significantly affect access to linker DNA. The absence of an effect of reducing temperatures on the rate of digestion of fibers, as compared to short oligonucleosomes, indicated that breathing motions to allow access to the fiber interior were not required for cleavage of linker DNA.
- Research Organization:
- California Univ., Los Angeles (USA)
- OSTI ID:
- 5209229
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHROMATIN
CLEAVAGE
CRYSTAL STRUCTURE
DNA
ENZYMES
ESTERASES
HYDROLASES
IMMOBILIZED ENZYMES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MICROCOCCUS
MICROORGANISMS
MICROSTRUCTURE
MOLECULAR STRUCTURE
NUCLEASES
NUCLEI
NUCLEIC ACIDS
NUCLEOSOMES
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
PHOSPHORUS 35
PHOSPHORUS ISOTOPES
RADIOISOTOPES
TRACER TECHNIQUES