Application Notes

The Decondensation factor 31 specifically interacts with histones H3 and H4 but not H2A and H2B

Issue link: https://resources.nanotempertech.com/i/1050563

Contents of this Issue

Navigation

Page 0 of 1

1 Characterization of a Protein - Histone Interaction Application Note NT-MO-015 The Decondensation factor 31 specifically interacts with histones H3 and H4 but not H2A and H2B Dr. Thomas Schubert Lab of Prof. Gernot Längst, University of Regensburg in cooperation with 2bind GmbH, Germany Abstract Histone proteins are associated with DNA and form a nucleoprotein structure termed chromatin. Eukaryotic genomes are packed into chromatin for storage and transport of the DNA. To regulate access to DNA for various essential processes such as transcription and translation, histone modifying, DNA modifying and chromatin binding proteins specifically interact with chromatin structures. Here we characterize the specific interaction of the Decondensation factor 31 (Df31) with core histones. Df31 was recently shown to form a complex with snoRNAs and chromatin, to generate accessible higher order structures of chromatin. Introduction Chromatin is the combination of DNA and proteins that make up the contents of the nucleus of a eukaryotic cell. Chromatin compacts the DNA, so it fits into the cell nucleus, helps to prevent DNA damage, and to control gene expression and other DNA dependent processes. The primary protein components of chromatin are histones that associate with DNA in 1.65 helical turns, forming an octamer of the histone proteins H2A, H2B, H3 and H4. The resulting structure, the nucleosome, is the basic structural component of chromatin (van-Holde, 1989). The histones within nucleosomes serve as docking platforms for modifying enzymes and other proteins. The modification of chromatin structure is essential for its function (Luger and Richmond, 1998; Wolffe, 1997). Chromatin modifications determine the degree of chromatin compaction and were shown to be associated with the differential regulation of the genomic target sites. In a recent study the Drosophila Decondensation factor 31 (Df31) was shown to be involved in regulating chromatin compaction and to be tethered to chromatin (Schubert et al., 2012). To identify the interaction partner of Df31 within the nucleosome, MicroScale Thermophoresis analyses were performed. Results In this application note, the binding behavior of Df31 to the recombinant core histones H2A, H2B, H3 and H4 was evaluated, using MicroScale Thermophoresis. Fig. 1 Measurement of Df31-EGFP binding to core histones. MST values were normalized to fraction bound. 3 independent measurements were performed to obtain the depicted binding curve. The K d of the Df31-H3 interaction was 1.5 ± 0.15 µM, the K d of the Df31-H4 interaction was 12 ± 0.6 µM.

Articles in this issue

view archives of Application Notes - The Decondensation factor 31 specifically interacts with histones H3 and H4 but not H2A and H2B