Although transmission of the gene expression program from mother to daughter cells has been suggested to be mediated by gene bookmarking, the precise mechanism by which bookmarking mediates post-mitotic transcriptional re-activation has been unclear. Here, we used a real-time gene expression system to quantitatively demonstrate that transcriptional activation of the same genetic locus occurs with a significantly more rapid kinetics in post-mitotic cells versus interphase cells. RNA polymerase II large subunit (Pol II) and bromodomain protein 4 (BRD4) were recruited to the locus in a different sequential order on interphase initiation versus post-mitotic re-activation resulting from the recognition by BRD4 of increased levels of histone H4 Lys 5 acetylation (H4K5ac) on the previously activated locus. BRD4 accelerated the dynamics of messenger RNA synthesis by de-compacting chromatin and hence facilitating transcriptional re-activation. Using a real-time quantitative approach, we identified differences in the kinetics of transcriptional activation between interphase and post-mitotic cells that are mediated by a chromatin-based epigenetic mechanism.
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We thank R. Singer (Albert Einstein College of Medicine, USA) for the MS2 clone, K. Ozato (NIH, USA) for the BRD4 clone, E. Verdin (UCSF, USA) for the BRD4-Δ1; Δ2; Δ1Δ2 constructs and C. D. Allis (The Rockfeller University, USA) for TH4 antibody. We greatly appreciate the gift of JQ1 from J. Bradner (Dana-Farber Cancer Institute, USA) and C. Vakoc. We thank C. Vakoc, A. A. Chakraborty and W. P. Tansey (Vanderbilt University, USA) for helpful comments and suggestions on ChIP assays. We also would like to thank Y. (S.) Mao, B. Zhang, M. S. Bodnar and M. R. Hübner, as well as other members of the Spector laboratory, for discussions and comments throughout the course of this work. This work was supported by a grant from NIH/NIGMS 42694, 42694-2OS1 (ARRA Supplement) to D.L.S.
The authors declare no competing financial interests.
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