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Sequencing of RNA and footprints of wildtype and RpS5b mutant Drosophila ovaries

ABSTRACT: Sequencing of RNA and footprints of wildtype and RpS5b mutant Drosophila ovaries

PROVIDER: PRJNA714404 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13961016.fastq.gz	Fastqsanger.gz
	SRR13961017.fastq.gz	Fastqsanger.gz
	SRR13961018.fastq.gz	Fastqsanger.gz
	SRR13961019.fastq.gz	Fastqsanger.gz
	SRR13961020.fastq.gz	Fastqsanger.gz

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Sequencing of RNA and footprints of wildtype and RpS5b mutant Drosophila ovaries [RIBO-seq]

Project description:Sequencing of RNA and footprints of wildtype and RpS5b mutant Drosophila ovaries [RIBO-seq]

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Sequencing of RNA and footprints of wildtype and RpS5b mutant Drosophila ovaries [RNA-seq]

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Project description:Purpose: To identify significantly different individual mRNA species or genetic pathways in the GCNAKO mutant sample when compared to the wildtype sample. GCNA is encoded by CG14814 (Flybase ID: FBgn0023515). Method: Total RNA was extracted using the Qiagen miRNEasy kit, followed by on column DNAse digest using the Qiagen DNAse digest kit. We used the miRNEasy kit to ensure collection of small RNA species as well as total RNA. RNA was collected from three independent bological replicates of freshly dissected Drosophila ovaries ( 100 ovaries per replicate), collected from 1-3 day old female Drosophila. Ovaries were flash frozen in liquid nitrogen after dissection to preserve the RNA species and prevent degradation. RNAse free conditions were maintained throughout the extraction protocol. RNA quality was tested by the sequencing core before sequencing the RNA.

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Sequencing of RNA and footprints of wildtype and RpS5b mutant Drosophila ovaries [RIBO-seq]

Project description:Emerging evidence suggests that ribosome heterogeneity may have important functional consequences in the translation of specific mRNAs within different cell types and under various conditions. Ribosome heterogeneity comes in many forms including post-translational modification of ribosome proteins (RPs), lack of specific RPs, and different RP paralogs incorporation. The Drosophila genome encodes for two RPS5 paralogs, RpS5a and RpS5b. While RpS5a is expressed ubiquitously, RpS5b exhibits enriched expression in the reproductive system. Deletion of RpS5b results in female sterility marked by developmental arrest of egg chambers at stages 7-8 and posterior follicle cell (PFC) hyperplasia. Both phenotypes are caused by defects in the germline and over-expression of RpS5a can compensate it. RpS5b mutant display increased rRNA transcription and RP production, accompanied by increased protein synthesis. Loss of RpS5b leads to microtubule-based defects and mislocalization of several factors, leading failure of Notch pathway activation in PFCs. Together, our results indicate that germ cell specific expression of RpS5b promotes proper egg chamber development by regulating both ribosome homeostasis and protein trafficking during mid-oogenesis.

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