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Determination of transcript's polyA tail length in Wild-Type and Mov10 KO mouse ESCs.

ABSTRACT: Determination of transcript's polyA tail length in Wild-Type and Mov10 KO mouse ESCs.

PROVIDER: PRJNA753146 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15390195_1.fastq.gz	Fastqsanger.gz
	SRR15390196_1.fastq.gz	Fastqsanger.gz
	SRR15390197_1.fastq.gz	Fastqsanger.gz
	SRR15390198_1.fastq.gz	Fastqsanger.gz

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Determination of transcript's polyA tail length in Wild-Type and Mov10 KO mouse ESCs.

Project description:The RNA helicase MOV10 targets mRNAs to promote their decay. MOV10 can directly bind to N6-methyladenosine (m6A) nucleotides, the most prevalent type of mRNA internal modification in mammals. Here we investigate the mechanism by which MOV10 promotes decay by looking at changes in the polyA tail of its targets. PolyA tail length shortening is usually associated with transcript decay. However, we did not observe changes in the polyA profile for MOV10 targets in MOV10-depleted and Wild-Type controls. The observation was true for targeted transcripts containing the m6A modification and for targeted transcripts without the modification. We conclude that MOV10-mediated decay does not perturb the polyA tail dynamics of its targets in ESCs.

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MOV10 knockout and gene expression

Project description:Transposable elements constitute nearly half of the mammalian genome and play important roles in genome evolution. While a multitude of both transcriptional and post-transcriptional mechanisms exist to silence transposable elements, control of transposition in vivo remains poorly understood. MOV10, an RNA helicase, is an inhibitor of mobilization of retrotransposons and retroviruses in cell culture assays. Here we report that MOV10 restricts LINE1 retrotransposition in mice. Although MOV10 is broadly expressed, its loss causes only incomplete penetrance of embryonic lethality, and the surviving MOV10-deficient mice are healthy and fertile. Biochemically, MOV10 forms a complex with UPF1, a key component of the nonsense-mediated mRNA decay pathway, and primarily binds to the 3' UTR of somatically expressed transcripts in testis. Consequently, loss of MOV10 results in an altered transcriptome in testis. Analyses using a LINE1 reporter transgene reveal that loss of MOV10 leads to increased LINE1 retrotransposition in somatic and reproductive tissues from both embryos and adult mice. Moreover, the degree of LINE1 retrotransposition inhibition is dependent on the Mov10 gene dosage. Furthermore, MOV10 deficiency reduces reproductive fitness over successive generations. Our findings demonstrate that MOV10 attenuates LINE1 retrotransposition in a dosage-dependent manner in mice.

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PolyA-tail enriched RNA-seq in differentiating TX1072 XX mESCs

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Identification of the cellular RNAs bound by MOV10

Project description:Using the iCLIP protocol we have identified the cellular RNA entities that are bound by MOV10. We report the location and sequence of the MOV10 binding region on each RNA entity. To identify the RNAs that bound MOV10, we UV-cross-linked HEK293F cells and immunoprecipitated with an irrelevant antibody (ir or "control") followed by a MOV10-specific antibody (MOV10) to isolate associated RNAs after stringent washing.

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FMRP-associated MOV10 facilitates and antagonizes miRNA-mediated regulation

Project description:The fragile X mental retardation protein FMRP is an RNA binding protein that regulates translation of its bound mRNAs through incompletely defined mechanisms. FMRP has been linked to the microRNA pathway and we show here that it is associated with MOV10, a putative helicase that is also associated with the microRNA pathway. We show that FMRP associates with MOV10 in an RNA-dependent manner and facilitates MOV10-association with RNAs in brain. We identified the RNA sequences recognized by MOV10 using iCLIP and found an increased number of G-quadruplexes in the CLIP sites. We provide evidence that MOV10 facilitates microRNA-mediated translation regulation and also has the novel role of increasing the expression of a subset of RNAs by sterically hindering Argonaute2 association. In summary, we have identified a new mechanism for FMRP-mediated translational regulation through its association with MOV10. Comparison of MOV10 siRNA knockdown, irrelevant siRNA control and MOV10 overexpression on total RNA levels

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2023-08-30 | PXD041814 | Pride

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