Mapping protein-DNA interactions in vivo with formaldehyde: Evidence that histone H4 is retained on a highly transcribed gene
Abstract
We have used formaldehyde-mediated protein-DNA crosslinking within intact cells to examine the in vivo chromatin structure of the D. melanogaster heat shock protein 70 (hsp70) genes. In agreement with previous in vitro studies, we find that the heat shock-mediated transcriptional induction of the hsp70 genes perturbs their chromatin structure, resulting in fewer proteinDNA contacts crosslinkable in vivo by formaldehyde. However, contrary to earlier in vitro evidence that histones may be absent from actively transcribed genes, we show directly, by immunoprecipitation of in vivo-crosslinked chromatin fragments, that at least histone H4 remains bound to hsp70 DNA in vivo, irrespective of its rate of transcription. The formaldehyde-based in vivo mapping techniques described in this work are generally applicable, and can be used both to probe proteinDNA interactions within specific genes and to determine the genomic location of specific chromosomal proteins.
Additional Information
© 1988 Cell Press. Received 4 December 1987, Revised 17 March 1988. We are greatly indebted to David Stollar and Arno Greenleaf for the anti-H4 and anti-RNA polymerase sera, respectively We also thank Mary Lou Pardue and Karen Travers for DNA clones and SL-2 cells, Darnel Finley, Bonnie Bartel, John McGrath, and Edward Winter for comments on the manuscript; and Barbara Doran for secretarial assistance. This work was supported by grants to A. V from the National Institutes of Health (CA43309 and GM33401) M J S was supported by a predoctoral fellowship from the National Science Foundation P. L. L. was supported by a postdoctoral fellowship from the American Cancer Society. The costs of publication of this article were defrayed m 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.Additional details
- Eprint ID
- 107824
- DOI
- 10.1016/s0092-8674(88)90469-2
- Resolver ID
- CaltechAUTHORS:20210129-142403364
- CA43309
- NIH
- GM33401
- NIH
- NSF Graduate Research Fellowship
- American Cancer Society
- Created
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2021-02-01Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field