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Comprehensive profiling of Roquin binding preferences for RNA stem-loops

ABSTRACT: Comprehensive profiling of Roquin binding preferences for RNA stem-loops

PROVIDER: PRJNA1128607 | ENA |

REPOSITORIES: ENA

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Comprehensive profiling of Roquin binding preferences for RNA stem-loops

Project description:Roquin proteins target specific RNA stem-loop motifs with their unique ROQ domain for mRNA degradation. The cellular levels of mRNAs are controlled by these post-transcriptional cis-regulatory stem-loop elements in the 3'-untranslated region (3'-UTR). Here, we first provide an RBNS (RNA Bind-n-Seq) experiment using pre-structured RNA libraries as input. This sRBNS (structured RBNS) was used to map Roquin's RNA binding preferences at nucleotide resolution, revealing preferences for loop sizes and consensus motifs for CDEs and ADEs.

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RNA-binding preference of the Roquin-1 CCCH-type zinc finger

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Roquin promotes constitutive mRNA decay via a conserved class of stem-loop recognition motifs.

Project description:The degradation of TNFM-NM-1 mRNA is critical for restricting TNFM-NM-1 synthesis and involves a constitutive decay element (CDE) in the 3M-bM-^@M-^Y untranslated region of the mRNA. We demonstrate that the active CDE is a stem-loop RNA motif that is recognized by the RNA-binding protein Roquin. Deep sequencing of Roquin-associated mRNAs identified CDE-containing mRNAs as the primary targets of Roquin on a transcriptome-wide scale. Expression profiles of LPS treated Raw264.7 cells (input), after Roquin IP or control IP were generated by deep sequencing, in three biological replicates, using Illumina HiSeq 2000.

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The RNA-binding proteins Roquin-1 and Regnase-1 interact and cooperate to prevent autoimmunity

Project description:This study aimed to analyze the transcriptomic changes occuring after knockout of alleles encoding for the RNA-binding proteins Roquin-1 and Roquin-2 (DKO), Regnase-1 (KO) or a combination of all three (TKO) in Th1 cells without or after restimulation with anti-CD3/28 antibodies. Data analysis revealed that in unstimulated as well as restimulated T cells of all three knockout genotypes a number of genes is differentially expressed compared to WT cells, with the strongest deregulation seen in TKO cells. A comparison of gene expression changes of all three genotypes indicated that cooperativity of Roquin and Regnase-1 is the prevalent mode of target regulation.

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Analysis of Roquin targetome during HCMV infection

Project description:To investigate how Roquin regulates cellular transcripts during Human cytomegalovirus (HCMV) infection, we examined the levels of cellular transcripts in cells treated control or Roquin-targeting siRNA during HCMV replication. Also, we performed Roquin crosslinking and immunoprecipitation followed by high-throughput sequencing (Roquin CLIP-seq) in HCMV-infected cells to identify which transcripts are directly bound by Roquin.

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DNA binding preferences of fungal SREBP transcription regulators

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