Labelling of Histone H5 and Its Interaction with DNA. 2. Cooperative Binding of Histone H5 to DNA as Probed By Steady-State Fluorescence and Diffusion-Enhanced Energy Transfer

Lerho, M.; Favazza, Marisa; Houssier, Claude

doi:10.1080/07391102.1990.10508567

Cited by 4 publications

(2 citation statements)

References 49 publications

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“…The fluorescence lifetimes were measured with an Edinburgh photo-counting instrument model 199S, equipped with a microsecond Xenon flash lamp model 199XF as reported elsewhere (47). Wensel et al (48) successfully applied this method to drug-DNAinteractions and showed that the difference between energy transfer rate constant~ o from TbHED3A o to DNA-bound (~b 0 ) and free (k 2 n ethidium is sufficient to allow an estimation of the accessibility of the terbium probe, free to diffuse in solution, with respect to each bound chromophore.…”

Section: Diffusion Enhanced Energy Transfer (Delet)mentioning

confidence: 99%

Interaction of Two Peptide-Acridine Conjugates Containing the SPKK Peptide Motif with DNA and Chromatin

Flock

Bailly

et al. 1994

Journal of Biomolecular Structure and Dynamics

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The interaction between DNA and two peptide-acridine conjugates containing one (1) or two (2) moieties of the Ser-Pro-Lys-Lys (SPKK) minor groove-binding peptide motif has been studied by a combination of hydrodynamic, biochemical and spectroscopic methods including diffusion-enhanced luminescence energy transfer (DELET) measurements with a Tb(III) lanthanide chelate as donor. Viscometric titrations do not reveal any significant difference between the two hybrid molecules which both unwind (by about 15 degrees) and extend the DNA similarly. DELET measurements show that the acridinyl chromophore of compounds 1 and 2 is much more accessible than that of a simple monointercalating drug such as acridine orange or ethidium. The accessibility factor increases proportionally with the peptide length, reflecting the extent of perturbation imposed upon the intercalating chromophore by the binding to DNA of the peptide moiety of the hybrids. Experiments with the osmium tetroxide-bispyridine reagent indicate that the two hybrid compounds both affect the local conformation of DNA rendering certain thymine residues conspicuously accessible to the probe. The drug-induced sites of hyperreactivity towards OsO4 in DNA are very similar with the exception of a short run of three T residues which is attacked more strongly in the presence of tetrapeptide-acridine conjugate 1 than with the octapeptide-acridine conjugate 2. These results are fully in agreement with previous footprinting studies and support the view that a minimum of two SPKK motifs is required to mimic the AT-specific minor groove binding antibiotic netropsin. On the basis of the DNA-binding properties of these two peptide-acridine hybrids, we present DNA-binding models in which the acridinyl moiety of compound 1 protrudes slightly outside the double helix but remains more or less parallel to the plane of the base-pairs. In contrast, with compound 2, where the octapeptide SPKKSPKK is bound to the minor groove, we postulate that the chromophore lies only partially overlapped with the base pairs in the intercalation site and, in addition, the heterocyclic chromophore is significantly tilted with respect to the double helix axis. Electric linear dichroism and DELET measurements with chromatin reveal that the presence of histone proteins affects the intercalative binding of compound 2 while it has practically no effect on the binding of compound 1.

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Section: Diffusion Enhanced Energy Transfer (Delet)mentioning

confidence: 99%

Interaction of Two Peptide-Acridine Conjugates Containing the SPKK Peptide Motif with DNA and Chromatin

Flock

Bailly

et al. 1994

Journal of Biomolecular Structure and Dynamics

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“…A H5(1-65), ~ H5(66-189), 0 H5(59-189) and • H5(32-185). Antisera were diluted 1:1000.Energy transfer from the neutral donor chelate ThHED3A to the acceptors was determined from changes in terbium fluorescence lifetime(19). Solutions for energy transfer measurements were prepared by addition ofThHED3A, at a final concentration of 1 mM, to the solutions of acceptor.…”

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confidence: 99%

Diffusion-Enhanced Energy Transfer Investigation of Histone H5 in Chromatin with a Fluorescently-Labelled Antibody Fragment Fab′

Sarlet

Muller²,

Houssier

1992

Journal of Biomolecular Structure and Dynamics

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The location of chicken erythrocyte H5 histone relative to the axis the 30 nm chromatin fibre axis has been investigated by diffusion-enhanced energy transfer. In this investigation, a neutral lanthanide chelate as donor and a fluorescent probe specific to H5 as acceptor have been used. The acceptor probe consists of H5 antibody Fab' fragment, which has been labeled with 5-iodoacetamidofluorescein (5-IAF). Using H5 fragments we have shown by ELISA that the antibodies recognized the N- and C-terminal ends of this histone. A neutral chelate of terbium (TbHED3A) was chosen as a suitable donor for energy transfer with IAF-labelled Fab' (Fab'-IAF) bound to H5 in various chromatin structures. The ionic strength dependence of the energy transfer from TbHED3A to chromatin-bound Fab'-IAF was used to estimate the accessibility and the location of the Fab' in chromatin. The rate constants for energy transfer, obtained from the lifetimes of the TbHED3A excited state in presence and absence of acceptor, indicated a decrease in transfer efficiency upon increase of salt concentration from 5 to 80 mM NaCl. This can be correlated with the chromatin folding occurring in this ionic strength range and is consistent with the location of at least some of the N and C-termini of H5 within the condensed chromatin structure.

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[13] Fluorescence resonance energy transfer

Selvin¹

1995

Methods in Enzymology

569

450

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Labelling of Histone H5 and Its Interaction with DNA. 2. Cooperative Binding of Histone H5 to DNA as Probed By Steady-State Fluorescence and Diffusion-Enhanced Energy Transfer

Cited by 4 publications

References 49 publications

Interaction of Two Peptide-Acridine Conjugates Containing the SPKK Peptide Motif with DNA and Chromatin

Interaction of Two Peptide-Acridine Conjugates Containing the SPKK Peptide Motif with DNA and Chromatin

Diffusion-Enhanced Energy Transfer Investigation of Histone H5 in Chromatin with a Fluorescently-Labelled Antibody Fragment Fab′

[13] Fluorescence resonance energy transfer

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