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The evolution of genic piRNA cluster loci expression patterns

ABSTRACT: The evolution of genic piRNA cluster loci expression patterns

PROVIDER: PRJNA264407 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR1617570.fastq.gz	Fastqsanger.gz
	SRR1617580.fastq.gz	Fastqsanger.gz
	SRR1617561.fastq.gz	Fastqsanger.gz
	SRR1617571.fastq.gz	Fastqsanger.gz
	SRR1617581.fastq.gz	Fastqsanger.gz

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The evolution of genic piRNA cluster loci expression patterns

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The evolution of genic piRNA cluster loci expression patterns

Project description:This study examines the conservation of Piwi-interacting RNA (piRNA) clusters that come from protein coding gene transcripts. By sequencing small RNA libraries from gonad tissues of Drosophilids and Glires we discover a diverse set of genic piRNA clusters conserved across animals. This dataset reveals new expression patterns for genic piRNA clusters and examines whether changing piRNA expression patterns correlates with sequence changes in piRNA cluster genomic sequence across a variety of animal species. We dissected gonad tissues consisting of ovaries from wildtype adult Drosophilids (D.melanogaster, D.erecta, D.yakuba, D.virilis) and testes from wildtype pre-pubsecent and adult mouse and rats. Adult testes from rabbits were purchased from Pel-Freez Biologicals. Total or immunopreciptated RNAs were extracted from the pulvirized gonad tissues of Drosophilids and Glires. Small RNAs were purified from these samples, converted into cDNA libraries, and sequenced on an Illumina HiSeq2000.

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Project description:In order to control transposable element (TE) activity, PIWI-interacting RNAs (piRNAs) have been evolved to silence TE transcriptionally and post-transcriptionally, and produced from heterochromatic genomic loci, called piRNA cluster. Maternal inherited piRNAs transmission is considered as the key step of piRNA cluster maintenance and induction of de nove piRNA cluster formation, however, how the original piRNAs were produced without maternal piRNAs deposition remains unclear. In this paper, we created a repetitive GFP reporter and found a Rhino-dependent piRNA cluster conversion spontaneously occurs to the reporter over generations. Importantly, this conversion is reversible by removal of maternal piRNAs of GFP reporter. Taking advantage of this reversible GFP reporter piRNA cluster, we found that siRNA is able to initiate piRNA cluster formation epigenetically, even when only maternal siRNA presence is capable to do so. Our results suggest the importance of siRNA on initiation of piRNA biogenesis and describe a whole picture of initiation and maintenance of piRNA cluster.

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Project description:Transposable element (TE) activity is repressed in animal gonads by PIWI-interacting RNAs (piRNAs) produced by piRNA clusters. Current models propose that piRNA clusters are functionally defined by the maternal inheritance of piRNAs produced during the previous generation. Taking advantage of an inactive cluster of P-element derived transgene insertions in Drosophila melanogaster, we show here that raising flies at high temperature (29°C) instead of 25°C results in stable conversion of this locus into an active piRNAs producing one reporting thus the first case of the establishment of an active piRNA cluster by environmental changes and without maternal inheritance of homologous piRNAs.

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The Cutoff protein regulates piRNA cluster expression and piRNA production in the Drosophila germline

Project description:In a broad range of organisms, Piwi-interacting RNAs (piRNAs) have emerged as core components of a surveillance system that protects the genome by silencing transposable and repetitive elements. A vast proportion of piRNAs is produced from discrete genomic loci, termed piRNA clusters. The molecular mechanisms and the factors that govern the expression of these loci are largely unknown. We have preciously shown the Cutoff (Cuff), a protein with similarity to yeast Rai1, is a component of the piRNA pathway. In order to understand the function of the Cuff protein in piRNA production, we produced small RNA libraries from cn, bw (wt) and cuffwm25 mutant ovaties. The analysis of these libraries revealed that approximately 80% of the total piRNA population is depleted in the absence of a functional Cuff protein. We also determined that Cuff is mostly a nuclear protein and is enriched at the level of certain piRNA clusters. Our results point to a role for Cuff in the transcriptional regulation of piRNA generating loci and in the production of the proper piRNA complement during Drosophila oogenesis. piRNA profiling in ovaries from cn, bw and cuffwm25 mutant ovaries

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