Preparation and structural characterization of nucleosomal core particles lacking one H2A.H2B dimer.
Nucleosomal core particles lacking one H2A.H2B dimer, (H2A.H2B)1 (H3.H4)2/DNA (146 bp), have been prepared by treatment of nucleosomal cores with dimethylmaleic anhydride, a reversible reagent for protein amino groups. The preparative procedure is simple, produces quantitative conversion of nucleosomal cores into dimer-deficient cores without formation of other subnucleosomal particles, and can be applied to the preparation of different H2A.H2B-deficient mono and oligonucleosomal particles. The structural properties of the dimer-deficient cores and complete nucleosomal cores reconstituted from the deficient particles and H2A.H2B dimers have been studied by DNase I digestion, thermal denaturation and circular dichroism.
de la Escalera
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Centro de Biología Molecular, Universidad Autónoma de Madrid, Cantoblanco, Spain.
Nieto
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MA
Palacián
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D000818:Animals; D002498:Centrifugation; D002645:Chickens; D002942:Circular Dichroism; D006657:Histones / Q000502:physiology*; D006358:Hot Temperature; D066298:In Vitro Techniques; D008299:Maleic Anhydrides / Q000494:pharmacology; D009707:Nucleosomes / Q000187:drug effects / Q000648:ultrastructure*
D006657:Histones; D008299:Maleic Anhydrides; D009707:Nucleosomes; C007474:2,3-dimethylmaleic anhydride