Elucidating the genetic and molecular basis of SWI/SNF-driven divergent non-coding transcription
(2017) KBKM01 20171Pure and Applied Biochemistry
- Abstract
- The discovery of pervasive transcription in eukaryotic organisms reveals that majority of promoters produce divergent non-coding transcripts besides their annotated transcripts. A recent study proposed the activation of divergent transcription by SWI/SNF chromatin remodelers. Mutating SNF5 subunit of the complex resulted in decreased levels of divergent transcription. Based on this finding, genomic replacement study was conducted. The yeast non-essential gene deletion library mutants were combined with snf5Δ. The screen aimed in identifying factors that suppress the snf5Δ and increase the levels of divergent non-coding transcription. SIN mutations in histones were reported to suppress the mutants of SWI/SNF complex. This hypothesis was... (More)
- The discovery of pervasive transcription in eukaryotic organisms reveals that majority of promoters produce divergent non-coding transcripts besides their annotated transcripts. A recent study proposed the activation of divergent transcription by SWI/SNF chromatin remodelers. Mutating SNF5 subunit of the complex resulted in decreased levels of divergent transcription. Based on this finding, genomic replacement study was conducted. The yeast non-essential gene deletion library mutants were combined with snf5Δ. The screen aimed in identifying factors that suppress the snf5Δ and increase the levels of divergent non-coding transcription. SIN mutations in histones were reported to suppress the mutants of SWI/SNF complex. This hypothesis was tested by introducing snf5Δ into two SIN mutants from yeast histone mutant library. To monitor the transcripts, a bidirectional promoter construct was introduced into the mutants. The bidirectional promoter drives two fluorescent proteins in the coding and non-coding direction respectively. The fluorescence read-outs from high-throughput flow cytometry analysis showed estimates of coding and non-coding transcripts. Screening for suppressor factors from the non-essential gene deletion library resulted in 21 potential candidate factors. Re-transformation of two candidates SPT21 and HDA1 into snf5Δ background resulted in ratio of mCherry/YFP (coding/non-coding) signal weakly suppressing the effect of snf5Δ. However, a SIN mutant H4 V43 had less ratio of signal compared to snf5Δ testing positive for the hypothesis. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/8908868
- author
- Gowthaman, Uthra LU
- supervisor
- organization
- course
- KBKM01 20171
- year
- 2017
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- eukaryotic organisms, SWI/SNF, transcripts, applied biochemistry
- language
- English
- id
- 8908868
- date added to LUP
- 2022-08-01 12:25:27
- date last changed
- 2022-08-01 12:25:27
@misc{8908868, abstract = {{The discovery of pervasive transcription in eukaryotic organisms reveals that majority of promoters produce divergent non-coding transcripts besides their annotated transcripts. A recent study proposed the activation of divergent transcription by SWI/SNF chromatin remodelers. Mutating SNF5 subunit of the complex resulted in decreased levels of divergent transcription. Based on this finding, genomic replacement study was conducted. The yeast non-essential gene deletion library mutants were combined with snf5Δ. The screen aimed in identifying factors that suppress the snf5Δ and increase the levels of divergent non-coding transcription. SIN mutations in histones were reported to suppress the mutants of SWI/SNF complex. This hypothesis was tested by introducing snf5Δ into two SIN mutants from yeast histone mutant library. To monitor the transcripts, a bidirectional promoter construct was introduced into the mutants. The bidirectional promoter drives two fluorescent proteins in the coding and non-coding direction respectively. The fluorescence read-outs from high-throughput flow cytometry analysis showed estimates of coding and non-coding transcripts. Screening for suppressor factors from the non-essential gene deletion library resulted in 21 potential candidate factors. Re-transformation of two candidates SPT21 and HDA1 into snf5Δ background resulted in ratio of mCherry/YFP (coding/non-coding) signal weakly suppressing the effect of snf5Δ. However, a SIN mutant H4 V43 had less ratio of signal compared to snf5Δ testing positive for the hypothesis.}}, author = {{Gowthaman, Uthra}}, language = {{eng}}, note = {{Student Paper}}, title = {{Elucidating the genetic and molecular basis of SWI/SNF-driven divergent non-coding transcription}}, year = {{2017}}, }