Abstract
The majority of DNA in eukaryotic cells is packaged by histones and many poorly characterized nonhistone proteins to form a dynamic structure known as chromatin. Chromatin is a periodic structure made up of repeating, regularly spaced subunits, the nucleosomes. Elegant genetic experiments have clearly demonstrated that histones play a central role in transcriptional control (1). Moreover, histones, via protein-protein interactions or by playing an architectural role, can facilitate or inhibit the transcriptional activation process (1,2). It also appears that the function of histones themselves may be regulated by protein modifications and therefore may be targets for cell signaling pathways (3,4).
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Grunstein, M., Hecht, A., Fisher-Adams, G., Wan, J., Mann, R. K., Strahl-Bolsinger S, Laroche, T., and Gasser, S. (1995) The regulation of euchromatin and heterochromatin by histones in yeast. J. Cell Sci. 19, 29–36.
Wolffe, A. P. (1994) Architectural transcription factors. Science 264, 1100,1101.
Wade, P. and Wolffe, A. P. (1997) Histone acetyltransferases in control. Curr. Biol. 7, 82–84.
Roth, S. Y. and Allis, C. D. (1996) Histone acetylation and chromatin assembly: a single escort, multiple dances? Cell 87, 5–8.
Workman, J. L. and Buchmann, A. R. (1993) Multiple functions of nucleosomes and regulatory functions in transcription. Trends Biochem. Sci. 18, 90–95.
Becker, P. B. (1994) The establishment of active promoters in chromatin. BioEssays 16, 541–547.
Lu, Q., Wallrath, L. L., and Elgin, S. C. R. (1994) Nucleosome positioning and gene regulation. J. Cell. Biochem. 55, 83–92.
Edmonson, D. G. and Roth, S. Y. (1996) Chromatin and transcription. FASEB J. 10, 1173–1182.
Wolffe, A. P. and Pruss, D. (1996) Deviant nucleosomes: the functional specialisation of chromatin. Trends Genetics 12, 58–62.
Tremethick, D. J. and Frommer, M. (1992) Partial purification, from Xenopus laevis oocytes, of an ATP-dependent activity required for nucleosome spacing in vitro. J. Biol. Chem. 267, 15,041–15,048.
Shimamura, A., Tremethick, D., and Worcel, A. (1988) Characterisation of the repressed 5S DNA minichromosomes assembled in vitro with a high speed supernatant of Xenopus laevis oocytes. Mol. Cell. Biol. 8, 4257–4269.
Almouzni, G., Mechali, M., and Wolffe, A. P. (1990) Competition between transcription complex assembly and chromatin assembly on replicating DNA. EMBO J. 9, 573–581.
Tsukiyama, T., Becker, P. B., and Wu, C. (1994) ATP-dependent nucleosome disruption at a heat-shock promoter mediated by binding of GAGA transcription factor. Nature 367, 525–532.
Kamakaka, R., Bulger, M., and Kadonaga, J. T. (1993) Potentiation of RNA polymerase II transcription by Gal 4-VP 16 during but not after DNA replication and chromatin assembly. Genes Dev. 7, 1779–1795.
Wolffe, A. P. (1991) Xenopus transcription factors: key molecules in the developmental regulation of differential gene expression. Biochem. J. 278, 313–324.
Glikin, G. C., Ruberti, I., and Worcel, A. (1984) Chromatin assembly in Xenopus oocytes: in vitro studies. Cell 37, 33–41.
Ito, T., Bulger, M., Pazin, M. J., Kobayashi, R., and Kadonaga, J. T. (1997) ACF, an ISWI-containing and ATP-utilising chromatin assembly and remodelling factor. Cell 90, 145–155.
Varga-Weisz, P. D., Wilm, M., Bonte, E., Dumas, K., and Becker, P. B. (1997) Chromatin-remodelling factor CHRAC contains the ATPases ISWI and topoisomerase II. Nature 388, 598–602.
Tremethick, D., Zucker, K., and Worcel, A. (1990) The transcription complex of the 5 S RNA gene but not Transcription Factor IIIA alone, prevents nucleosomal repression of transcription. J. Biol. Chem. 265, 5014–5023.
Kleinschmidt, J. A. and Seiter, A. (1988) Soluble acidic complexes containing histones H3 and H4 in nuclei in Xenopus laevis oocytes. Cell 29, 799–809.
Ng, K. W., Ridgway, P., Cohen, D. R., and Tremethick, D. J. (1997) The binding of a fos/jun heterodimer can completely disrupt the structure of a nucleosome. EMBO J. 16, 2072–2085.
Dignam, J. D., and Lebovitz, R. M., and Roeder, R. G. (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 11, 1475–1489.
Drew, H. R. and Calladine, C. R. (1987) Sequence-specific positioning of core histones on an 860 base-pair DNA. J. Mol. Biol. 195, 143–173.
Tremethick, D. J. (1994) High mobility group proteins 14 and 17 can space nucleosomal particles deficient in histones H2A and H2B creating a template that is transcriptional active. J. Biol. Chem. 269, 28,436–28,442.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc.
About this protocol
Cite this protocol
Tremethick, D.J. (1999). Preparation of Chromatin Assembly Extracts from Xenopus Oocytes. In: Becker, P.B. (eds) Chromatin Protocols. Methods in Molecular Biology™, vol 119. Humana Press. https://doi.org/10.1385/1-59259-681-9:175
Download citation
DOI: https://doi.org/10.1385/1-59259-681-9:175
Publisher Name: Humana Press
Print ISBN: 978-0-89603-665-9
Online ISBN: 978-1-59259-681-2
eBook Packages: Springer Protocols