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  • 99
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    Spontaneous DNA unwrapping from the 5S nucleosome (nuc) is stimulated two- to fivefold upon removal of the core histone tail domains. Nucleosomes reconstituted with the 215-bp 5S DNA fragment were selected to eliminate minor translational positions, incubated with trypsin agarose beads to remove core histone tail domains, and digested with either <t>EcoRV</t> or <t>BbvI.</t> A 154-bp truncated 5S DNA fragment was included in each digest as a naked DNA reference. (A) Proteins within intact or tailless nucleosomes were examined on SDS-18% PAGE. Lane 1, (H3/H4) 2 ). (C) Intact or tailless 5S nucleosomes and 154-bp DNA fragments were digested with the indicated enzymes, and the logarithm of the uncut fraction was plotted versus digestion times. Note that the nucleosomal curves were generated from digests containing 5,000 or 600 U of EcoRV or BbvI, respectively, while the naked DNA curves were generated from identical reaction mixtures containing 10-U/ml concentrations of each enzyme. Diamonds, intact nucleosome; squares, tailless nucleosome; and triangles, naked DNA. (C) K conf eq . W.t. and tryp, intact and tailless nucleosomes, respectively.
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    Spontaneous DNA unwrapping from the 5S nucleosome (nuc) is stimulated two- to fivefold upon removal of the core histone tail domains. Nucleosomes reconstituted with the 215-bp 5S DNA fragment were selected to eliminate minor translational positions, incubated with trypsin agarose beads to remove core histone tail domains, and digested with either <t>EcoRV</t> or <t>BbvI.</t> A 154-bp truncated 5S DNA fragment was included in each digest as a naked DNA reference. (A) Proteins within intact or tailless nucleosomes were examined on SDS-18% PAGE. Lane 1, (H3/H4) 2 ). (C) Intact or tailless 5S nucleosomes and 154-bp DNA fragments were digested with the indicated enzymes, and the logarithm of the uncut fraction was plotted versus digestion times. Note that the nucleosomal curves were generated from digests containing 5,000 or 600 U of EcoRV or BbvI, respectively, while the naked DNA curves were generated from identical reaction mixtures containing 10-U/ml concentrations of each enzyme. Diamonds, intact nucleosome; squares, tailless nucleosome; and triangles, naked DNA. (C) K conf eq . W.t. and tryp, intact and tailless nucleosomes, respectively.
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    Spontaneous DNA unwrapping from the 5S nucleosome (nuc) is stimulated two- to fivefold upon removal of the core histone tail domains. Nucleosomes reconstituted with the 215-bp 5S DNA fragment were selected to eliminate minor translational positions, incubated with trypsin agarose beads to remove core histone tail domains, and digested with either <t>EcoRV</t> or <t>BbvI.</t> A 154-bp truncated 5S DNA fragment was included in each digest as a naked DNA reference. (A) Proteins within intact or tailless nucleosomes were examined on SDS-18% PAGE. Lane 1, (H3/H4) 2 ). (C) Intact or tailless 5S nucleosomes and 154-bp DNA fragments were digested with the indicated enzymes, and the logarithm of the uncut fraction was plotted versus digestion times. Note that the nucleosomal curves were generated from digests containing 5,000 or 600 U of EcoRV or BbvI, respectively, while the naked DNA curves were generated from identical reaction mixtures containing 10-U/ml concentrations of each enzyme. Diamonds, intact nucleosome; squares, tailless nucleosome; and triangles, naked DNA. (C) K conf eq . W.t. and tryp, intact and tailless nucleosomes, respectively.
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    Spontaneous DNA unwrapping from the 5S nucleosome (nuc) is stimulated two- to fivefold upon removal of the core histone tail domains. Nucleosomes reconstituted with the 215-bp 5S DNA fragment were selected to eliminate minor translational positions, incubated with trypsin agarose beads to remove core histone tail domains, and digested with either <t>EcoRV</t> or <t>BbvI.</t> A 154-bp truncated 5S DNA fragment was included in each digest as a naked DNA reference. (A) Proteins within intact or tailless nucleosomes were examined on SDS-18% PAGE. Lane 1, (H3/H4) 2 ). (C) Intact or tailless 5S nucleosomes and 154-bp DNA fragments were digested with the indicated enzymes, and the logarithm of the uncut fraction was plotted versus digestion times. Note that the nucleosomal curves were generated from digests containing 5,000 or 600 U of EcoRV or BbvI, respectively, while the naked DNA curves were generated from identical reaction mixtures containing 10-U/ml concentrations of each enzyme. Diamonds, intact nucleosome; squares, tailless nucleosome; and triangles, naked DNA. (C) K conf eq . W.t. and tryp, intact and tailless nucleosomes, respectively.
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    Image Search Results


    Spontaneous DNA unwrapping from the 5S nucleosome (nuc) is stimulated two- to fivefold upon removal of the core histone tail domains. Nucleosomes reconstituted with the 215-bp 5S DNA fragment were selected to eliminate minor translational positions, incubated with trypsin agarose beads to remove core histone tail domains, and digested with either EcoRV or BbvI. A 154-bp truncated 5S DNA fragment was included in each digest as a naked DNA reference. (A) Proteins within intact or tailless nucleosomes were examined on SDS-18% PAGE. Lane 1, (H3/H4) 2 ). (C) Intact or tailless 5S nucleosomes and 154-bp DNA fragments were digested with the indicated enzymes, and the logarithm of the uncut fraction was plotted versus digestion times. Note that the nucleosomal curves were generated from digests containing 5,000 or 600 U of EcoRV or BbvI, respectively, while the naked DNA curves were generated from identical reaction mixtures containing 10-U/ml concentrations of each enzyme. Diamonds, intact nucleosome; squares, tailless nucleosome; and triangles, naked DNA. (C) K conf eq . W.t. and tryp, intact and tailless nucleosomes, respectively.

    Journal: Molecular and Cellular Biology

    Article Title: The Core Histone N-Terminal Tail Domains Negatively Regulate Binding of Transcription Factor IIIA to a Nucleosome Containing a 5S RNA Gene via a Novel Mechanism

    doi: 10.1128/MCB.25.1.241-249.2005

    Figure Lengend Snippet: Spontaneous DNA unwrapping from the 5S nucleosome (nuc) is stimulated two- to fivefold upon removal of the core histone tail domains. Nucleosomes reconstituted with the 215-bp 5S DNA fragment were selected to eliminate minor translational positions, incubated with trypsin agarose beads to remove core histone tail domains, and digested with either EcoRV or BbvI. A 154-bp truncated 5S DNA fragment was included in each digest as a naked DNA reference. (A) Proteins within intact or tailless nucleosomes were examined on SDS-18% PAGE. Lane 1, (H3/H4) 2 ). (C) Intact or tailless 5S nucleosomes and 154-bp DNA fragments were digested with the indicated enzymes, and the logarithm of the uncut fraction was plotted versus digestion times. Note that the nucleosomal curves were generated from digests containing 5,000 or 600 U of EcoRV or BbvI, respectively, while the naked DNA curves were generated from identical reaction mixtures containing 10-U/ml concentrations of each enzyme. Diamonds, intact nucleosome; squares, tailless nucleosome; and triangles, naked DNA. (C) K conf eq . W.t. and tryp, intact and tailless nucleosomes, respectively.

    Article Snippet: Intact or tailless 5S nucleosomes and the 154-bp DNA fragment were incubated together with EcoRV (5,000 U/ml or 10 U/ml) or BbvI (600 U/ml or 10 U/ml) in the manufacturer's recommended buffer (New England BioLabs) for the times indicated in the legend to Fig. .

    Techniques: Incubation, Polyacrylamide Gel Electrophoresis, Generated