Premium
Characterization of the electrophoretic properties of nucleosome core particles by transverse polyacrylamide pore gradient gel electrophoresis
Author(s) -
Orbán Lászlo,
Garner Mark M.,
Wheeler David,
Tietz Dietmar,
Chrambach Andreas
Publication year - 1993
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501401114
Subject(s) - nucleosome , dna , electrophoresis , gel electrophoresis , gel electrophoresis of nucleic acids , biophysics , radius , chemistry , bent molecular geometry , crystallography , chromatography , biology , biochemistry , histone , computer security , organic chemistry , computer science
Transverse pore gradient gel electrophoresis, previously applied to bent DNA, has extended the usefulness of the gel retardation assay in two ways: (i) by differentiating between different DNA conformations; (ii) by providing information regarding the physical properties of DNA. In the present study, similarly extended information is obtained with regard to a well‐characterized DNA‐protein complex, the chicken erythrocyte nucleosome core particle. (i) The winding of DNA around the protein core constrains the DNA which renders its Ferguson curve (migration distance vs. gel concentration) similar to that of kinetoplast DNA, i.e. it intersects sharply with the Ferguson curves of linear DNA standards. By contrast, the deproteinized nucleosome DNA exhibits a Ferguson curve similar to linear standards of the same lenght. (ii) Interpretation of the Ferguson curve based on a mathematical model shows that the nucleosome exhibits a linear Ferguson plot [log(mobility) vs. gel concentration]. This is similar to and characteristic of spherical proteins, contrasting with the concave plot typical for linear and bent DNA. (iii) The effective size of the nucleosome, evaluated in terms of an “equivalent sphere” ( i.e. a hypothetical spherical particle with a radius, R es , having the same electrophoretic mobility as DNA for a particular set of experimental conditions), remains invariant across the gel concentration range of 3–9 % T. This is similar to proteins and bacteriophages and contrasts with the progressive decline of R es with increasing gel concentration observed for linear DNA and the deproteinized nucleosomal DNA.