TY - JOUR
T1 - Mapping proteinDNA interactions in vivo with formaldehyde
T2 - Evidence that histone H4 is retained on a highly transcribed gene
AU - Solomon, Mark J.
AU - Larsen, Pamela L.
AU - Varshavsky, Alexander
N1 - Funding Information:
We are greatly indebted to David Stollar and Arno Greenleaf for the ant=-H4 and antt-RNA polymerase sera, respect=vely We also thank Mary Lou Pardue and Karen Travers for DNA clones and SL-2 cells, Darnel Finley, Bonme 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 fellowshtp from the American Cancer Society.
PY - 1988/6/17
Y1 - 1988/6/17
N2 - We have used formaldehyde-mediated proteinDNA 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 shockmediated 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 vivocrosslinked 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.
AB - We have used formaldehyde-mediated proteinDNA 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 shockmediated 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 vivocrosslinked 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.
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U2 - 10.1016/S0092-8674(88)90469-2
DO - 10.1016/S0092-8674(88)90469-2
M3 - Article
C2 - 2454748
AN - SCOPUS:0023948705
SN - 0092-8674
VL - 53
SP - 937
EP - 947
JO - Cell
JF - Cell
IS - 6
ER -