title: Transcriptional elongation requires DNA break-induced signalling.
abstract: We have previously shown that RNA polymerase II (Pol II) pause release and transcriptional elongation involve phosphorylation of the factor TRIM28 by the DNA damage response (DDR) kinases ATM and DNA-PK. Here we report a significant role for DNA breaks and DDR signalling in the mechanisms of transcriptional elongation in stimulus-inducible genes in humans. Our data show the enrichment of TRIM28 and γH2AX on serum-induced genes and the important function of DNA-PK for Pol II pause release and transcriptional activation-coupled DDR signalling on these genes. γH2AX accumulation decreases when P-TEFb is inhibited, confirming that DDR signalling results from transcriptional elongation. In addition, transcriptional elongation-coupled DDR signalling involves topoisomerase II because inhibiting this enzyme interferes with Pol II pause release and γH2AX accumulation. Our findings propose that DDR signalling is required for effective Pol II pause release and transcriptional elongation through a novel mechanism involving TRIM28, DNA-PK and topoisomerase II.
authors.affiliation: Department of Radiation Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Heeyoun
authors.identifier: http://orcid.org/0000-0003-0038-1985
authors.initials: H
authors.lastname: Bunch
authors.affiliation: Center for Cancer Computational Biology, Dana Farber Cancer Institute, Boston, Massachusetts 02130, USA.
authors.forename: Brian P
authors.initials: BP
authors.lastname: Lawney
authors.affiliation: Department of Radiation Oncology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
authors.forename: Yu-Fen
authors.initials: YF
authors.lastname: Lin
authors.affiliation: Department of Radiation Oncology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
authors.forename: Aroumougame
authors.initials: A
authors.lastname: Asaithamby
authors.affiliation: Department of Radiation Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Ayesha
authors.initials: A
authors.lastname: Murshid
authors.affiliation: Center for Cancer Computational Biology, Dana Farber Cancer Institute, Boston, Massachusetts 02130, USA.
authors.forename: Yaoyu E
authors.initials: YE
authors.lastname: Wang
authors.affiliation: Department of Radiation Oncology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
authors.forename: Benjamin P C
authors.initials: BP
authors.lastname: Chen
authors.affiliation: Department of Radiation Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Stuart K
authors.identifier: http://orcid.org/0000-0001-9966-3354
authors.initials: SK
authors.lastname: Calderwood
mesh_terms: D047369:Chromatin Immunoprecipitation; D020552:Comet Assay; D053960:DNA Breaks; D004249:DNA Damage / Q000235:genetics; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D051747:DNA-Activated Protein Kinase / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D057809:HEK293 Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D018613:Microscopy, Confocal; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D042925:Positive Transcriptional Elongation Factor B / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D012097:Repressor Proteins / Q000378:metabolism*; D015398:Signal Transduction; D061805:Transcription Elongation, Genetic; D014158:Transcription, Genetic / Q000235:genetics; D000076183:Tripartite Motif-Containing Protein 28
chemical_list: D006657:Histones; D009687:Nuclear Proteins; D012097:Repressor Proteins; C101116:TRIM28 protein, human; D000076183:Tripartite Motif-Containing Protein 28; D042925:Positive Transcriptional Elongation Factor B; D051747:DNA-Activated Protein Kinase; C497263:PRKDC protein, human; D012319:RNA Polymerase II; D004250:DNA Topoisomerases, Type II