CREB-binding Protein (CBP)/p300 and RNA Polymerase II Colocalize in Transcriptionally Active Domains in the Nucleus

doi:10.1083/jcb.150.1.265

Published online 11 July 2000. doi:10.1083/jcb.150.1.265

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© The Rockefeller University Press, 0021-9525/2000/7/265/ $5.00
The Journal of Cell Biology, Volume 150, Number 1, July 10, 2000 265-274

Reports

CREB-binding Protein (CBP)/p300 and RNA Polymerase II Colocalize in Transcriptionally Active Domains in the Nucleus

Anna von Mikecz^a,e, Suisheng Zhang^b, Marc Montminy^d, Eng M. Tan^e, and Peter Hemmerich^c,e
^a Junior Research Group of Molecular Cell Biology, Medizinisches Institut für Umwelthygiene, Heinrich-Heine-Universät Düsseldorf, 40225 Düsseldorf, Germany
^b Department of Biochemistry, Institut für Molekulare Biotechnologie, 07745 Jena, Germany
^c Department of Molecular Biology, Institut für Molekulare Biotechnologie, 07745 Jena, Germany
^d The Salk Institute, La Jolla, California 92037
^e Department of Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037

Correspondence to: Peter Hemmerich, Department of Molecular Biology, Institut für Molekulare Biotechnologie, Beutenbergstr. 11, 07745 Jena, Germany. Tel:49-3641-656262 Fax:49-3641-656225 E-mail:phemmer{at}imb-jena.de.

The spatial organization of transcription- associated proteinsis an important control mechanism of eukaryotic gene expression.Here we analyzed the nuclear distribution of the transcriptionalcoactivators CREB-binding protein (CBP)/p300 in situ by confocallaser scanning microscopy, and in vivo complex formation bycoimmunoprecipitation. A subpopulation of CBP and p300 is targetedto active sites of transcription and partially colocalizes withhyper- and hypophosphorylated RNA polymerase II (pol II) indiscrete regions of variable size throughout the nucleus. However,the coactivators were found in tight association with hypophosphorylated,but not hyperphosphorylated pol II. Transcriptional inhibitioninduced a relocation of CBP/p300 and pol II into speckles. Moreover,double and triple immunofluorescence analyses revealed the presenceof CBP, p300, and pol II in a subset of promyelocytic leukemia(PML) bodies. Our results provide evidence for a dynamic spaciallink between coactivators of transcription and the basal transcriptionmachinery in discrete nuclear domains dependent upon the transcriptionalactivity of the cell. The identification of pol II in CBP/PML-containingnuclear bodies supports the idea that transcription takes placeat PML bodies.

Key Words: transcription, coactivators, nuclear body, RNA polymerase II,promyelocytic leukemia

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