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The crystal structure of the NHL domain in complex with RNA reveals the molecular basis of Drosophila Brain tumor-mediated gene regulation

ABSTRACT: Here, we analyze the RNA-binding preferences of several NHL containing RNA-binding proteins (BRAT, NCL-1, TRIM71, WECH, LIN-41, TRIM56, and MEI-P26) using RNAcompete.

ORGANISM(S): synthetic construct

PROVIDER: GSE73000 | GEO | 2015/09/14

SECONDARY ACCESSION(S): PRJNA295968

REPOSITORIES: GEO

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The crystal structure of the NHL domain in complex with RNA reveals the molecular basis of Drosophila Brain tumor-mediated gene regulation

Project description:Here, we analyze the RNA-binding preferences of several NHL containing RNA-binding proteins (BRAT, NCL-1, TRIM71, WECH, LIN-41, TRIM56, and MEI-P26) using RNAcompete. In the RNAcompete assay, purified GST-tagged RNA-binding protein is incubated with an excess of RNA pool and bound RNA from individual pulldown experiments are directly labeled and hybridized to a custom Agilent 244K microarray.

2015-09-14 | E-GEOD-73000 | biostudies-arrayexpress

Simultaneous activation of Tor and suppression of ribosome biogenesis by TRIM-NHL proteins promotes terminal differentiation

Project description:Tissue development and homeostasis depend on the balance between growth and terminal differentiation, but the mechanisms coordinating these processes remain elusive. Accumulating evidence indicates that ribosome biogenesis (RiBi) and protein synthesis, two cellular processes sustaining growth, are tightly regulated and yet can be uncoupled during stem cell differentiation. Using the Drosophila adult female germline stem cell (GSC) and larval neuroblast (NB) systems, we show that Mei-P26 and Brat, two Drosophila TRIM-NHL paralogs, are responsible for uncoupling RiBi and protein synthesis during differentiation. In differentiating cells, Mei-P26 and Brat activate the Target of rapamycin (Tor) kinase to promote translation, while concomitantly repressing RiBi. Depletion of Mei-P26 or Brat results in defective terminal differentiation, which can be rescued by ectopic activation of Tor together with suppression of RiBi. Our results indicate that uncoupling RiBi and translation activities by TRIM-NHL activity creates the conditions required for terminal differentiation.

2023-03-03 | GSE218205 | GEO

Identification of Meiotic-P26-bound RNAs in cutlured Drosophila SL2 cells by iCLIP (individual-nucleotide resolution UV crosslinking and immunoprecipitation)

Project description:The TRIM-NHL protein Meiotic P26 (Mei-P26) governs gene expression and controls stem cell fate in Drosophila. Mei-P26 represses the translation of the key regulator protein Nanos and induces miRNA-dependent silencing to regulate stem cell renewal, differentiation and proliferation. The structural and mechanistic details of mRNA target recognition and RNA binding by its C-terminal NHL domain remain elusive. To identify its cellular target RNAs, we performed iCLIP2 experiments in cultured Drosophila SL2 cells that were transfected with Flag-tagged Mei-P26 full-length, or its NHL domain.

2021-06-01 | GSE152013 | GEO

Genome-wide identification of BRAT- and PUM-associated mRNAs reveals PUM-independent functions for BRAT in translational repression and mRNA degradation during the Drosophila maternal-to-zygotic transition

Project description:Here, we analyze the RNA-binding preferences of the Drosophila melanogaster BRAT protein using RNAcompete. In the RNAcompete assay, purified GST-tagged BRAT is incubated with an excess of RNA pool and bound RNA from individual pulldown experiments are directly labeled and hybridized to a custom Agilent 244K microarray.

2015-03-30 | E-GEOD-60498 | biostudies-arrayexpress

Project description:Here, we analyze the RNA-binding preferences of the Drosophila melanogaster BRAT protein using RNAcompete.

2015-03-30 | GSE60498 | GEO

The RNA hairpin binder TRIM71 modulates alternative splicing by repressing Mbnl1

Project description:We demonstrate that global effects of TRIM71 on the cellular transcriptome of mouse embryonic stem cells (mESCs) are largely attributable to its RNA binding activity, and map the transcriptome-wide targets of wild-type TRIM71, NHL domain mutant TRIM71, and RING domain mutant TRIM71.

2019-08-01 | GSE130248 | GEO

microRNA expression analysis in fly ovaries from wuho and mei-p26 mutants

Project description:Purpose: The goal of this study is to analyze microRNA affected by wuho and mei-p26 mutations in Drosophila ovary Ovarian microRNA prolife of 7 day old control flies and flies bearing wuho and mei-p26 mutations, in duplicate.Sample libraries were prepared using the QIAseq miRNA Library Kit (QIAGEN) according to the manufacturer's protocols. Adaptors are ligated sequentially to the 3' and 5' ends of miRNAs. Subsequently, universal cDNA synthesis with UMI assignment, cDNA cleanup, library amplification and library cleanup are performed. Libraries were sequenced on an Illumina instrument (75-cycle single-end read, 75SE). Sequencing data was processed using the Illumina software BCL2FASTQ v2.20.

2020-01-03 | GSE142830 | GEO

Profiling of Brat associated mRNAs from Drosophila embryos by RIP-CHIP

Project description:The Drosophila TRIM-NHL protein Brain tumor (Brat) plays important roles during early embryogenesis, in cell fate decisions, during neurogenesis and in mature neurons. Brat is an RNA-binding protein and functions as translational repressor. However, which RNAs Brat regulates and how RNA-binding specificity is achieved, is unknown. Using RNA-Immunoprecipitation we identify Brat-bound mRNAs in Drosophila embryos and define a consensus binding motif. Microarrays were used to profile Brat-associated RNAs from fly embryos.

2015-11-13 | GSE71663 | GEO

The tumor suppressor Brat controls neuronal lineages by inhibiting the transcription factors Deadpan and Zelda

Project description:The TRIM-NHL protein Brain tumor (Brat) acts as a tumor suppressor in the Drosophila brain, but how it suppresses tumor formation is not completely understood. Here, we combine temperature controlled brat RNAi with transcriptome analysis to identify the immediate brat targets in Drosophila neuroblasts. Besides the known target Deadpan (Dpn), our experiments identify the transcription factor Zelda (Zld) as a critical target of brat. Our data show that Zld is expressed in neuroblasts and required to allow re-expression of Dpn in transit amplifying intermediate neural progenitors. Upon neuroblast division, Brat is enriched in one daughter cell where its NHL domain directly binds to specific motifs in the 3’UTR of dpn and zld mRNA to mediate their degradation. In brat mutants, both Dpn and Zld continue to be expressed, but inhibition of either transcription factor prevents tumorigenesis. Our genetic and biochemical data indicate that Dpn inhibition requires higher Brat levels than Zld inhibition and suggest a model where stepwise post-transcriptional inhibition of distinct factors ensures sequential generation of fates in a stem cell lineage.

2017-11-28 | GSE104592 | GEO

Transcription profiling of human non-Hodgkins lymphoma (NHL) cell lines

Project description:We profiled 40 NHL cell lines to determine gene expression patterns and molecular subtypes. Experiment Overall Design: RNA profiles of NHL cell lines.

2009-05-16 | E-GEOD-15329 | biostudies-arrayexpress

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