1. Transcriptional elongation requires DNA break-induced signalling
- Author
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Ayesha Murshid, Aroumougame Asaithamby, Brian Lawney, Yaoyu E. Wang, Stuart K. Calderwood, Heeyoun Bunch, Yu Fen Lin, and Benjamin P C Chen
- Subjects
Chromatin Immunoprecipitation ,Transcription Elongation, Genetic ,TRIM28 ,Transcription, Genetic ,Positive Transcriptional Elongation Factor B ,DNA damage ,Fluorescent Antibody Technique ,General Physics and Astronomy ,Repressor ,RNA polymerase II ,DNA-Activated Protein Kinase ,Tripartite Motif-Containing Protein 28 ,Real-Time Polymerase Chain Reaction ,Article ,General Biochemistry, Genetics and Molecular Biology ,Histones ,03 medical and health sciences ,0302 clinical medicine ,Transcription (biology) ,Humans ,Phosphorylation ,030304 developmental biology ,0303 health sciences ,Microscopy, Confocal ,Multidisciplinary ,biology ,DNA Breaks ,Nuclear Proteins ,General Chemistry ,Molecular biology ,Cell biology ,Repressor Proteins ,body regions ,DNA Topoisomerases, Type II ,HEK293 Cells ,Histone ,biology.protein ,Comet Assay ,RNA Polymerase II ,Chromatin immunoprecipitation ,030217 neurology & neurosurgery ,DNA Damage ,Signal Transduction - 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., RNA polymerase II (Pol II) pause release and transcriptional elongation involve phosphorylation of TRIM28 by the DNA damage response (DDR) kinases. Here, Bunch et al. show that DDR signalling is coupled with and required for transcriptional elongation in stimulus-inducible genes and involves topoisomerase II.
- Published
- 2015
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