Nuclear magnetic resonance and biochemical studies of histone mobility in nucleosomes

Thesis/Dissertation · Tue Jan 01 04:00:00 EST 1985

OSTI ID:6985760

Smith, II, R M

/sup 13/C NMR spectra were recorded for nucleosome cores and nucleosome cores with specific segments removed in an effort to identify regions of histone mobility in the nucleosome core. Three types of particles were generated by limited proteolysis of nucleosome cores. Brief ..cap alpha..-chmyotrypsin treatment yielded a particle in which H3 residues 1-20 had been removed. Further ..cap alpha..-chymotrypsin treatment of this particle yielded a particle in which H3 residues 1-20, H2B residues 1-18, and H4 residues 1022 were absent. Very brief digestion of this particle with trypsin produced a particle in which H3 residues 1-20, H2B residues 1-18, H4 residues 1-22, and H2A residues 1-11 and 119-128 were removed. Comparisons of /sup 13/C NMR spectra of proteolysed nucleosome core particles with /sup 13/ C NMR data of native nucleosome cores and the compositions of different peptide regions with potential mobility showed that the random coil regions observed in the core particles were located in H3 residues 1-23 and H2A residues 1-16 and 116-128. Particles with the N-terminal tails removed still retained sedimentation and circular dichroism characteristics similar to intact core particles. Random coil resonances were also seen in preparations of nucleosome oligomers, nucleosome oligomers depleted of H1 and H5, nucleosome oligomers in a condensed form, and in intact nuclei. These resonances appeared to be very similar to those appearing in the nucleosome core spectrum.

• Research Organization: Florida State Univ., Tallahassee (USA)
• OSTI ID: 6985760
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BIOCHEMISTRY
CARBON 13
CARBON ISOTOPES
CHEMICAL REACTIONS
CHEMISTRY
CHROMATIN
CONFORMATIONAL CHANGES
DECOMPOSITION
ENZYMATIC HYDROLYSIS
EVEN-ODD NUCLEI
HISTONES
HYDROLYSIS
ISOTOPES
LIGHT NUCLEI
LYSIS
MAGNETIC RESONANCE
NUCLEAR MAGNETIC RESONANCE
NUCLEI
NUCLEOPROTEINS
NUCLEOSOMES
ORGANIC COMPOUNDS
PROTEINS
RESONANCE
SOLVOLYSIS
STABLE ISOTOPES