Acidic Patch Mutant H2BE105A,E113A Recombinant Nucleosome with Linker DNA, Biotinylated

The acidic patch is a negatively charged region of the nucleosome surface that serves as a conserved interaction hub for
neighboring nucleosomes and nucleosome binding proteins, often via salt bridges with arginine anchors [1]. The acidic patch
plays a critical role in chromatin condensation and chromatin remodeling [1-3]. Recombinant mononucleosomes containing
acidic patch mutations can be used to study the biological functions of the acidic patch.

H2BE105A,E113A Recombinant Nucleosome with Linker DNA consists of 199 base pairs of DNA wrapped around an
octamer core of histone proteins (two each of H2A, H2B, H3.1, and H4) to form a nucleosome, the basic repeating unit of
chromatin. The 5’ biotin-TEG DNA consists of a core 147 bp 601 nucleosome assembly sequence [4] flanked by 26 bp linker
sequences as underlined below. Histone H2B contains a glutamate-to-alanine (E-to-A) substitution at positions 105 and 113
(H2BE105A, E113A). H2BE105 and H2BE113 both reside in H2B alphaC helix extension and are associated with
nucleosome binding factors such as H4-N-terminal tail, LANA, RCC1, and HMGN2 [1]. H2BE105A,E113A disrupts binding
with SMARCB1/BAF47, a subunit of the SWI/SNF (BAF) family of chromatin remodeling complexes. These complexes serve a
critical role in cell division, cell and tissue differentiation, and development, and SWI/SNF complex malfunction has been linked
to over 20% of human cancers [5].