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RIP-seq of mouse Asterix/Gtsf1 expressed in Sf9 cells

ABSTRACT: RIP-seq of mouse Asterix/Gtsf1 expressed in Sf9 cells

PROVIDER: PRJNA634753 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11837410.fastq.gz	Fastqsanger.gz
	SRR11837411.fastq.gz	Fastqsanger.gz
	SRR11837412.fastq.gz	Fastqsanger.gz
	SRR11837413.fastq.gz	Fastqsanger.gz
	SRR11837414.fastq.gz	Fastqsanger.gz

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RIP-seq of mouse Asterix/Gtsf1 expressed in Sf9 cells

Project description:Asterix/Gtsf1 has been previously been shown to be involved in the silencing of retrotransposons in animal germ cells through the piRNA pathway. During early stages of purification of mouse Gtsf1 protein in Sf9 cells for structural studies, we found that the protein was co-purifying with endogenous nucleic acids. We first characterized those RNAs by next-generation sequencing (this dataset) and then later investigated its RNA-binding in more relevant cell culture systems. In each case, we found Asterix/Gtsf1 was binding tRNAs. Given the role of tRNAs as primers for Long Terminal Repeat (LTR) transposons, this work implicates Asterix/Gtsf1 as exploiting tRNA dependence to identify transposon transcripts and promote piRNA silencing.

2021-02-20 | GSE151110 | GEO

eCLIP of mouse Asterix/Gtsf1 transfected in mouse P19 embryonal teratoma cells

Project description:Asterix/Gtsf1 has been previously been shown to be involved in the silencing of retrotransposons in animal germ cells through the piRNA pathway. Using eCLIP and a custom informatic pipeline, we found that Asterix/Gtsf1 binds directly to RNAs and preferentially interacts with tRNAs as a class. Given the role of tRNAs as primers for Long Terminal Repeat (LTR) transposons, this work implicates Asterix/Gtsf1 as exploiting tRNA dependence to identify transposon transcripts and promote piRNA silencing.

2021-02-20 | GSE151108 | GEO

eCLIP of Drosophila Asterix/Gtsf1 transfected in Drosophila ovarian somatic sheath (OSS) cells

2021-02-20 | GSE151107 | GEO

eCLIP of Drosophila Asterix/Gtsf1 (endogenous promoter) in Drosophila ovarian somatic sheath (OSS) cells

2021-02-20 | GSE151109 | GEO

Asterix/Gtsf1 links tRNAs and piRNA silencing of retrotransposons

Project description:Asterix/Gtsf1 links tRNAs and piRNA silencing of retrotransposons

| PRJNA634754 | ENA

eCLIP of mouse Asterix/Gtsf1 transfected in mouse P19 embryonal teratoma cells

Project description:eCLIP of mouse Asterix/Gtsf1 transfected in mouse P19 embryonal teratoma cells

| PRJNA634751 | ENA

eCLIP of Drosophila Asterix/Gtsf1 transfected in Drosophila ovarian somatic sheath (OSS) cells

Project description:eCLIP of Drosophila Asterix/Gtsf1 transfected in Drosophila ovarian somatic sheath (OSS) cells

| PRJNA634750 | ENA

eCLIP of Drosophila Asterix/Gtsf1 (endogenous promoter) in Drosophila ovarian somatic sheath (OSS) cells

Project description:eCLIP of Drosophila Asterix/Gtsf1 (endogenous promoter) in Drosophila ovarian somatic sheath (OSS) cells

| PRJNA634752 | ENA

Sf9 induction with phenobarbital

Project description:Effect of phenobarbital on Sf9 cell cultures genes expression. RNA from phenobarbital treated Sf9 cell cultures were compared to control treated (DMSO) Sf9 cell

2018-09-22 | GSE32337 | GEO

Asterix/Gtsf1 links tRNAs and piRNA silencing of retrotransposons.

Project description:The Piwi-interacting RNA (piRNA) pathway safeguards genomic integrity by silencing transposable elements (transposons) in the germline. While Piwi is the central piRNA factor, others including Asterix/Gtsf1 have also been demonstrated to be critical for effective silencing. Here, using enhanced crosslinking and immunoprecipitation (eCLIP) with a custom informatic pipeline, we show that Asterix/Gtsf1 specifically binds tRNAs in cellular contexts. We determined the structure of mouse Gtsf1 by NMR spectroscopy and identified the RNA-binding interface on the protein's first zinc finger, which was corroborated by biochemical analysis as well as cryo-EM structures of Gtsf1 in complex with co-purifying tRNA. Consistent with the known dependence of long terminal repeat (LTR) retrotransposons on tRNA primers, we demonstrate that LTR retrotransposons are, in fact, preferentially de-repressed in Asterix mutants. Together, these findings link Asterix/Gtsf1, tRNAs, and LTR retrotransposon silencing and suggest that Asterix exploits tRNA dependence to identify transposon transcripts and promote piRNA silencing.

| S-EPMC8283736 | biostudies-literature

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