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Analysis of archaeal histone interactions from M. fervidus .
Author(s) -
Stencel Katherine Ann,
Tims Hannah S.
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.489.2
Subject(s) - histone , archaea , nucleosome , biology , förster resonance energy transfer , dna , plasmid , biochemistry , genetics , gene , fluorescence , physics , quantum mechanics
Archaea are prokaryotes with cellular characteristics of eukaryotes. Homologues of eukaryotic histones are found in some archaea, however, the structural and functional characteristics of archaeal nucleosomes are not well understood. Archaea can have 0‐6 histones, which are able to form hetero/ homodimers. Histones from the archaea, Methanothermus fervidus (HMfA, HMfB) were expressed and purified from E. coli plasmids containing clones of the histones (gift of Kathleen Sandman, The Ohio State University). Cloned archaeal high affinity DNA sequences were purified from E. coli to be used to assemble nucleosomes. The histones were tagged with fluorescent dyes to be used in Fluorescence resonance energy transfer (FRET) experiments to study histone‐histone and histone‐DNA interactions. To allow specific placement of the dyes, the histones were mutated to introduce cystine residues as attachment sites. Kathleen Sandman (OSU) provided mutated HMfB. HMfA will be mutated with a QuickChange mutagenesis kit (Stratagene) for hetero‐histone complex investigation. This work was supported by a grant to The College of Wooster from the Howard Hughes Medical Institute through the Undergraduate Science Education Program.