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Download StatisticsGrisan, Valentina (2019). Regulation of the transcription cycle by co-ordinate interaction of ATP-dependent chromatin remodeling and histone post-translational modifications. University of Birmingham. M.Sc.
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Grisan2019MScbyRes.pdf
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Abstract
Changes in chromatin structure are instrumental in controlling transcription initiation. This process is regulated by enzymes known as histone post-translational modifications (HPTMs).
This work investigates how the interaction between Nucleosome remodelling factor (NURF) and HPTMs regulates transcription.
Previous studies showed that NURF interacts with several HPTMs (H4K16Ac, H3K4me3, H3K9Ac and H3S10p) and it is responsible for histone octamer sliding.
ChIP-Seq was used to profile the distribution of HPTMs and NURF in both Drosophila S2 cells and primary hemocytes, showing these marks are capable of targeting NURF remodelling to the +1 nucleosome during transcription in vivo.
Specific anti-MRG15 antibodies were generated for Chip-Seq analysis using wild-type and Nurf301 mutant cells. We speculate that targeting of NURF to active genes re-establishes the correct spacing between nucleosomes which in normal conditions allows binding of MRG15 to H3K36me3 activating the Set2/Rpd3 pathway, known for preventing cryptic initiation.
mRNA-Seq and CAGE-Seq were performed to determine trascriptome profiles, indicating NURF does not modulate initiation site, rather transcription levels from individual genes.
We generated a Nurf301 dsRNAi S2 cell line and developed whole larval homogenisation procedure using Drosophila third instar larvae to isolate primary cells and further investigate the effect of loss of NURF on RNA polymerase initiation and elongation.
| Type of Work: | Thesis (Masters by Research > M.Sc.) | |||||||||
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| Award Type: | Masters by Research > M.Sc. | |||||||||
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| Licence: | All rights reserved | |||||||||
| College/Faculty: | Colleges (2008 onwards) > College of Medical & Dental Sciences | |||||||||
| School or Department: | Institute of Cancer and Genomic Sciences | |||||||||
| Funders: | Biotechnology and Biological Sciences Research Council | |||||||||
| Subjects: | Q Science > QH Natural history > QH301 Biology | |||||||||
| URI: | http://etheses.bham.ac.uk/id/eprint/9247 |
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