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Partial purification of the template-active fraction of chromatin: A preliminary report

Gottesfeld, J. M. and Garrard, W. T. and Bagi, G. and Wilson, R. F. and Bonner, James (1974) Partial purification of the template-active fraction of chromatin: A preliminary report. Proceedings of the National Academy of Sciences of the United States of America, 71 (6). pp. 2193-2197. ISSN 0027-8424.

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A fraction of rat-liver chromatin that is transcriptionally active in vivo has been purified 6- to 7-fold over whole chromatin. This was accomplished by selectively shearing chromatin with DNase II followed by fractionating the released portion on the basis of its solubility properties in 2 mM MgCl2. The resulting soluble material comprises 11% of the total chromatin DNA and is impoverished in histone and enriched in nonhistone protein. Compared with unsheared chromatin, this minor fraction exhibits marked differences in chromosomal protein species. DNA renaturation studies indicate that this fraction is composed of a specific subset of whole genomal DNA sequences. Furthermore, DNA·RNA hybridization experiments suggest that almost 60% of the nonrepetitious DNA sequences of this minor fraction could code for cellular RNA.

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Additional Information:© 1974 by the National Academy of Sciences. Contributed by James Bonner, March 18, 1974. We thank Dr. A. Douvas and Mssrs. R. Watson and M. Wilkes for assistance with this project. We also thank Drs. D. Holmes and B. Hough and Mr. W. Pearson for many helpful discussions and for critical evaluation of the manuscript. This work was supported by the U.S. Public Health Service (Grants GM 86 and GM 13762). W.T.G. was a recipient of a Damon Runyon Fund for Cancer Research Fellowship (DRF-755).
Subject Keywords:chromatin fractionation; DNase II; DNA•RNA hybridization
Issue or Number:6
Record Number:CaltechAUTHORS:GOTpnas74
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:8669
Deposited By: Tony Diaz
Deposited On:04 Sep 2007
Last Modified:02 Oct 2019 23:53

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