ISSN:
1433-4909
Keywords:
Key words Histone fold
;
Recombinant protein
;
Thermostability
;
DNA packaging
;
Archaeoglobus fulgidus
;
Circular dichroism
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract All archaeal histones studied to date have similar lengths, 66 to 69 amino acid residues that form three α-helices separated by two β-strand loop regions which together constitute a histone fold. In contrast, the eukaryal nucleosome core histones are larger, 102 to 135 residues in length, with N-terminal and C-terminal extensions flanking the histone fold that participate in gene regulation and higher-order chromatin assembly. In the Methanococcus jannaschii genome, MJ1647 was annotated as an open reading frame predicted to encode an archaeal histone with an approximately 27-amino-acid C-terminal extension, and we here document the DNA binding and assembly properties and thermodynamic stability parameters of the recombinant product of MJ1647 synthesized in Escherichia coli with (rMJ1647) and without (rMJ1647Δ) the C-terminal extension. The presence of the C-terminal extension did not prevent homodimer formation or inhibit DNA binding, but the complexes formed by rMJ1647, presumably archaeal nucleosomes containing a (rMJ1647)4 tetramer, were apparently less stable than those formed by (rMJ1647Δ)4. The presence of the C-terminal extension increased the thermostability of rMJ1647 when compared with rMJ1647Δ in 0.2 M KCl at pH 4 but not in the absence of KCl at pH 1. Based on thermal unfolding transitions, rMJ1647 and rHAfB generated by expression of AF0337 cloned from the genome of the related hyperthermophile Archaeoglobus fulgidus in E. coli were found to have higher thermodynamic stabilities than all previously studied archaeal histones.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s007920050006
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