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Saccharomyces cerevisiae

ABSTRACT: In vitro transcription start sites (TSSs) determined by 5'-RACE

PROVIDER: PRJNA361496 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5179254_1.fastq.gz	Fastqsanger.gz
	SRR5179254_2.fastq.gz	Fastqsanger.gz
	SRR5179255_1.fastq.gz	Fastqsanger.gz
	SRR5179255_2.fastq.gz	Fastqsanger.gz
	SRR5179256_1.fastq.gz	Fastqsanger.gz

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Project description:Both, acetylation of histones and of histone variant H2A.Z are conserved features of eukaryotic transcription start sites (TSSs) and both features appear to be critical for correct transcription initiation. However, complex patterns of transcriptional regulation have complicated the establishment of functional links between histone acetylation, H2A.Z deposition and their importance in transcription regulation. To elucidate these links, we took advantage of the unusual genome organization in Trypanosoma brucei, a highly divergent eukaryote. In T. brucei genes are organized in long polycistronic transcription units, drastically reducing the sites of transcription initiation. Employing a highly sensitive and quantitative mass-spectrometry-based approach, we quantified the genome-wide histone acetylation and methylation pattern and identified various acetyl and methyl marks exclusively enriched at TSSs In addition, we show that deletion of histone acetyltransferase 2 results in a loss of H4 acetylation at TSSs, a loss of H2A.Z deposition at TSSs and a shift in the sites of transcription initiation. Combined, our findings demonstrate an evolutionary conserved link between histone H4 acetylation, H2A.Z deposition and RNA transcription initiation.

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Global identification of transcription start sites in the genome of Apis mellifera by using 5’ LongSAGE

Project description:The 5’ LongSAGE (5’LS) approach provides a powerful genomic tool for identifying Transcription start sites (TSSs) in sequenced genome. The main purpose of this study is to identify the actual TSSs of expressed genes as well as the usage of different TSSs in Apis mellifera. We also wish to provide expression evidence for the in silico predicted genes and reveal some previously undiscovered genes. The transcriptome of adult drone head was fathomed through 5’LS analysis, and TSSs of the expressed genes were determined by mapping the 5’LS tag sequences to the honeybee genome sequences.

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Project description:The initial step of RNA polymerase II (Pol II) transcription involves a large number of transcription factors and arises at multiple sites within most promoters. TFIIH is an essential, multi-subunit transcription factor that assembles on promoter DNA with Pol II and five other general transcription factors (GTFs) to form a pre-initiation complex (PIC) for basal transcription. During transcription initiation, TFIIH melts promoter DNA through the ATPase activity of its Ssl2 subunit. In the model eukaryote Saccharomyces cerevisiae, after DNA melting, Pol II scans downstream for usable transcription start sites (TSSs). To understand the function of Ssl2/TFIIH in promoter scanning and TSS selection, we identified novel alleles of SSL2 in genetic screens for mutants defective in TSS distribution that may potentially arise from altered scanning. Consistent with this notion, these ssl2 alleles alter scanning in ways that are distinct from how changes to the Pol II active site alter scanning and this difference is observed genome-wide. Our investigations support two major pathways in controlling promoter scanning and TSS selection, one controlling the efficiency of initiation through Pol II activity or factors regulating Pol II activity; another network appears to control the processivity of scanning by Ssl2/TFIIH.

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