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RNA Interacts with Topoisomerase I to Modulate its Catalytic Activity

ABSTRACT: RNA Interacts with Topoisomerase I to Modulate its Catalytic Activity

PROVIDER: PRJNA880811 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29418917.fastq.gz	Fastqsanger.gz
	SRR29418918.fastq.gz	Fastqsanger.gz
	SRR29418919.fastq.gz	Fastqsanger.gz
	SRR29418920.fastq.gz	Fastqsanger.gz
	SRR29413833_1.fastq.gz	Fastqsanger.gz

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RNA Interacts with Topoisomerase I to Modulate its Catalytic Activity

Project description:Topoisomerase I (TOP1) is an essential enzyme that relaxes DNA to prevent and dissipate torsional stress during transcription. However, the mechanisms underlying the regulation of TOP1 activity remain elusive. Using eCLIP and UV-RIP-seq we identify TOP1 as a RNA binding protein (RBP). Using a TOP1 RNA binding mutant and TOP1cc-seq to map TOP1 catalytic activity, we reveal that RNA opposes TOP1 activity as RNAPII commences transcription of active genes. These findings support new insights into the coordinated actions of RNA and TOP1 in regulating DNA topological stress intrinsic to RNAPII-dependent transcription.

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Topoisomerase 3B Interacts with RNAi Machinery to Promote Heterochromatin Formation and Transcriptional Silencing

Project description:Topoisomerases solve topological problems during DNA metabolism, but whether they participate in RNA metabolism remains unclear. Top3b represents a new family of topoisomerases carrying activities for both DNA and RNA. Here we show that in Drosophila, Top3b interacts biochemically and genetically with the RNAi-induced silencing complex (RISC) containing AGO2, p68 RNA helicase, and FMRP. Top3b and RISC mutants are defective in heterochromatin formation and transcriptional silencing by position-effect variegation assay; and this defect is suppressed in their double mutants. Moreover, both Top3b and AGO2 single mutants exhibit reduced heterochromatin protein HP1 in pericentric heterochromatin; and this reduction is also suppressed in their double mutant. Furthermore, expression of several genes and transposable elements (TEs) in heterochromatin is increased in the Top3b mutant. Notably, Top3b mutants defective in either RNA binding or catalytic activity are deficient in promoting HP1 recruitment and silencing of TEs. Our data suggest that Top3b acts as an RNA topoisomerase in siRNA-guided heterochromatin formation and transcriptional silencing. Grant title: An RNA topoisomerase complex interacts with Fragile X syndrome protein to promote neurodevelopment and maintain normal life-span. Grant ID: AG000689-08. Source: NIA/NIH

2018-08-29 | GSE119185 | GEO

A dual-activity topoisomerase complex regulates mRNA translation and turnover

Project description:Topoisomerase 3β (TOP3B) and TDRD3 form a dual-activity topoisomerase complex that interacts with FMRP and can change the topology of both DNA and RNA. Here, we investigated the post-transcriptional influence of TOP3B and associated proteins on mRNA translation and turnover. First, we discovered that in human HCT116 colon cancer cells, knock-out (KO) of TOP3B had similar effects on mRNA turnover and translation as did TDRD3-KO, while FMRP-KO resulted in rather distinct effects, indicating that TOP3B had stronger coordination with TDRD3 than FMRP in mRNA regulation. Second, we identified TOP3B-bound mRNAs in HCT116 cells; we found that while TOP3B did not directly influence the stability or translation of most TOP3B target mRNAs, it stabilized a subset of target mRNAs but had a more complex effect on translation--enhancing for some mRNAs whereas reducing for others. Interestingly, a point mutation that specifically disrupted TOP3B catalytic activity only partially recapitulated the effects of TOP3B-KO on mRNA stability and translation, suggesting that the impact of TOP3B on target mRNAs is only in part linked to its ability to change topology of mRNAs. Collectively, our data suggest that TOP3B-TDRD3 can regulate mRNA translation and turnover by mechanisms that are dependent and independent of topoisomerase activity.

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Topoisomerase 3β Interacts with RNAi Machinery to Promote Heterochromatin Formation and Transcriptional Silencing in Drosophila [ChIP-Seq]

Project description:Topoisomerases solve topological problems during DNA metabolism, but whether they participate in RNA metabolism remains unclear. Top3b represents a family of topoisomerases carrying activities for both DNA and RNA. Here we show that in Drosophila, Top3b interacts biochemically and genetically with the RNAi-induced silencing complex (RISC) containing AGO2, p68 RNA helicase, and FMRP. Top3b and RISC mutants are similarly defective in heterochromatin formation and transcriptional silencing by position-effect variegation assay. Moreover, both Top3b and AGO2 mutants exhibit reduced levels of heterochromatin protein HP1 in pericentric heterochromatin. Furthermore, expression of several genes and transposable elements in heterochromatin is increased in the Top3b mutant. Notably, Top3b mutants defective in either RNA binding or catalytic activity are deficient in promoting HP1 recruitment and silencing of transposable elements. Our data suggest that Top3b may act as an RNA topoisomerase in siRNA-guided heterochromatin formation and transcriptional silencing.

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O-GlcNAcylation elevates topoisomerase IIα catalytic activity: a role in the chemotherapy resistance of breast cancer

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