Growth-Phase-Dependent Synthesis of Histones in the Archaeon Methanothermus fervidus

Histone preparations from Methanothermus fervidus (HMf) contain two small polypeptides, HMfA and HMfB, which in solution are dimers and compact DNA to form nucleosome-like structures. These archaeal nucleosome-like structures constrain positive DNA supercoils, in contrast to the negatively supercoil...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1994-12, Vol.91 (26), p.12624-12628
Main Authors: Sandman, Kathleen, Grayling, Rowan A., Dobrinski, Beate, Lurz, Rudi, Reeve, John N.
Format: Article
Language:English
Subjects:
Amino acids
Archaea - genetics
Archaea - growth & development
Bacterial Proteins - metabolism
Base Composition
Base Sequence
Biochemistry
Cell growth
Circular Dichroism
Deoxyribonucleic acid
DNA
DNA Primers - chemistry
DNA, Bacterial - genetics
DNA-Binding Proteins - metabolism
Electrophoresis
Gels
Genomes
Geometric growth
Histones
Histones - metabolism
Hot Temperature
Mass ratio
Methanothermus fervidus
Microscopy, Electron
Molecular Sequence Data
Molecules
Nucleic Acid Denaturation
Proteins
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Summary:Histone preparations from Methanothermus fervidus (HMf) contain two small polypeptides, HMfA and HMfB, which in solution are dimers and compact DNA to form nucleosome-like structures. These archaeal nucleosome-like structures constrain positive DNA supercoils, in contrast to the negatively supercoiled DNA in eukaryal nucleosomes. HMfA has been found to make up as much as 80% of HMf preparations synthesized by M. fervidus cells during the exponential growth phase of batch cultures but to decrease to ≈50% as cultures enter the stationary phase. By using a nondenaturing polyacrylamide gel system at pH 6.1, we have demonstrated that HMf preparations contain HMfA homodimers, HMfB homodimers, and HMfA-HMfB heterodimers and that heating a mixture of recombinant HMfA and HMfB homodimers at 95⚬C for 5 min generates HMfA-HMfB heterodimers. Circular dichroism spectroscopy indicates that HMfA and HMfB have very similar secondary structures, but based on agarose gel electrophoretic mobility shifts, DNA topology assays, and electron microscopy, they have different DNA binding properties. HMfA binding to DNA could be detected at lower protein/DNA ratios than HMfB, but HMfB binding resulted in more extensive DNA compaction. The increased HMfB synthesized in cells approaching the stationary phase and the highly compacted state of HMfB-bound DNA are consistent with preparations for the impending period of limited genome activity.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.91.26.12624