Nucleosome structural changes induced by binding of non-histone chromosomal proteins HMGN1 and HMGN2
| Autor: | Hideto Shimahara, Sailaja S. Seeram, Shin-ichi Tate, Takaaki Hirano, Kouichi Ohya, Shun Matsuta |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
HMGN
Unmodified recombinant histones SDS–PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis Solenoid (DNA) Article General Biochemistry Genetics and Molecular Biology Reconstitution PCR polymerase chain reaction RP-HPLC reverse phase high performance liquid chromatography Nucleosome Histone octamer NMR nuclear magnetic resonance CD circular dichroism biology IPTG isopropyl-β-d-galactopyranoside phH2A phH2B phH3 and phH4 vectors for the gene expression of all four recombinant human core histones H2A H2B H3 and H4 respectively Molecular biology Linker DNA SWI/SNF CD Histone LB Luria–Bertani Chromatosome biology.protein Biophysics HMG high mobility group HMGN1 HMGN2 non-histone chromosomal proteins MNase micrococcal nuclease |
| Zdroj: | FEBS Open Bio |
| ISSN: | 2211-5463 |
| DOI: | 10.1016/j.fob.2013.03.002 |
| Popis: | Interactions between the nucleosome and the non-histone chromosomal proteins (HMGN1 and HMGN2) were studied by circular dichroism (CD) spectroscopy to elucidate structural changes in the nucleosome induced by HMGN binding. Unlike previous studies that used a nucleosome extracted from living cells, in this study we utilized a nucleosome reconstituted from unmodified recombinant histones synthesized in Escherichia coli and a 189-bp synthetic DNA fragment harboring a nucleosome positioning sequence. This DNA fragment consists of 5′-TATAAACGCC-3′ repeats that has a high affinity to the histone octamer. A nucleosome containing a unique octamer-binding sequence at a specific location on the DNA was produced at sufficiently high yield for spectroscopic analysis. CD data have indicated that both HMGN1 and HMGN2 can increase the winding angle of the nucleosome DNA, but the extent of the structural changes induced by these proteins differs significantly. This suggests HMGN1 and HMGN2 would have different abilities to facilitate nucleosome remodeling. Highlights • A nucleosome was reconstituted from recombinant histones and a synthetic DNA. • Nucleosomes were produced at sufficiently high yield for spectroscopic analysis. • A nucleosome with and without HMGN proteins was analyzed using CD spectroscopy. • CD data indicate that HMGN proteins increase the winding angle of the nucleosome DNA. • HMGN1 and HMGN2 may have different abilities to facilitate nucleosome remodeling. |
| Databáze: | OpenAIRE |
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