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Analysis of Histones in Xenopus laevis: I. A DISTINCT INDEX OF ENRICHED VARIANTS AND MODIFICATIONS EXISTS IN EACH CELL TYPE AND IS REMODELED DURING DEVELOPMENTAL TRANSITIONS

Shechter, David ; Nicklay, Joshua J ; Chitta, Raghu K ; Shabanowitz, Jeffrey ; Hunt, Donald F ; Allis, C. David

The Journal of biological chemistry, 2009-01, Vol.284 (2), p.1064-1074 [Periódico revisado por pares]

United States: American Society for Biochemistry and Molecular Biology

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  • Título:
    Analysis of Histones in Xenopus laevis: I. A DISTINCT INDEX OF ENRICHED VARIANTS AND MODIFICATIONS EXISTS IN EACH CELL TYPE AND IS REMODELED DURING DEVELOPMENTAL TRANSITIONS
  • Autor: Shechter, David ; Nicklay, Joshua J ; Chitta, Raghu K ; Shabanowitz, Jeffrey ; Hunt, Donald F ; Allis, C. David
  • Assuntos: Alternative Splicing - genetics ; Animals ; Female ; Histones - classification ; Histones - genetics ; Histones - isolation & purification ; Histones - metabolism ; Male ; Protein Processing, Post-Translational ; Tissue Culture Techniques ; Transcription, Chromatin, and Epigenetics ; Xenopus laevis - embryology ; Xenopus laevis - genetics ; Xenopus laevis - growth & development ; Xenopus laevis - metabolism
  • É parte de: The Journal of biological chemistry, 2009-01, Vol.284 (2), p.1064-1074
  • Notas: ObjectType-Article-1
    SourceType-Scholarly Journals-1
    ObjectType-Feature-2
    content type line 23
    The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1–3.
    To whom correspondence should be addressed: Box 78, 1230 York Ave., New York, NY 10065. Tel.: 212-327-7839; E-mail: alliscd@rockefeller.edu.
    This work was supported, in whole or in part, by National Institutes of Health National Research Service Award/Kirschstein Fellowship GM075486 (to D. S.), Grant GM37537 (to D. F. H.), and Grant GM53512 (to C. D. A.). This work was also supported by Rockefeller University. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
  • Descrição: Histone proteins contain epigenetic information that is encoded both in the relative abundance of core histones and variants and particularly in the post-translational modification of these proteins. We determined the presence of such variants and covalent modifications in seven tissue types of the anuran Xenopus laevis, including oocyte, egg, sperm, early embryo equivalent (pronuclei incubated in egg extract), S3 neurula cells, A6 kidney cells, and erythrocytes. We first developed a new robust method for isolating the stored, predeposition histones from oocytes and eggs via chromatography on heparin-Sepharose, whereas we isolated chromatinized histones via conventional acid extraction. We identified two previously unknown H1 isoforms (H1fx and H1B.Sp) present on sperm chromatin. We immunoblotted this global collection of histones with many specific post-translational modification antibodies, including antibodies against methylated histone H3 on Lys⁴, Lys⁹, Lys²⁷, Lys⁷⁹, Arg², Arg¹⁷, and Arg²⁶; methylated histone H4 on Lys²⁰; methylated H2A and H4 on Arg³; acetylated H4 on Lys⁵, Lys⁸, Lys¹², and Lys¹⁶ and H3 on Lys⁹ and Lys¹⁴; and phosphorylated H3 on Ser¹⁰ and H2A/H4 on Ser¹. Furthermore, we subjected a subset of these histones to two-dimensional gel analysis and subsequent immunoblotting and mass spectrometry to determine the global remodeling of histone modifications that occurs as development proceeds. Overall, our observations suggest that each metazoan cell type may have a unique histone modification signature correlated with its differentiation status.
  • Editor: United States: American Society for Biochemistry and Molecular Biology
  • Idioma: Inglês
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