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Non-RNA binding domains of Upstream of N-Ras mediate RNA tertiary structure specificity

ABSTRACT: This study presents a novel paradigm of how multi-RBD proteins can achieve RNA target specificity by selecting certain RNA tertiary structures and provides a basis for reconstituting an entire RNP for future structure determination efforts.

PROVIDER: PRJEB37467 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR4352340.fastq.gz	Fastqsanger.gz
	ERR4352341.fastq.gz	Fastqsanger.gz
	ERR4352342.fastq.gz	Fastqsanger.gz
	ERR4352343.fastq.gz	Fastqsanger.gz
	ERR4352344.fastq.gz	Fastqsanger.gz

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Project description:Autoimmune diseases disproportionately affect females more than males. The XX sex chromosome complement is strongly associated with susceptibility to autoimmunity. Xist long noncoding RNA (lncRNA) is expressed only in females to randomly inactivate one of the two X chromosomes to achieve gene dosage compensation. Here, we show that the Xist ribonucleoprotein (RNP) complex, comprised of numerous autoantigenic components, is an important driver of sex-biased autoimmunity. Inducible transgenic expression of a non-silencing form of Xist in male mice introduced Xist RNP complexes and sufficed to produce autoantibodies. Male SJL/J mice expressing transgenic Xist developed more severe multiorgan pathology in pristane-induced model of lupus than wild-type males. Xist expression in males reprogrammed T and B cell population and chromatin states to more resemble wild type females. Human patients with autoimmune diseases displayed significant autoantibodies to multiple components of XIST RNP. Thus, a sex-specific lncRNA scaffolds ubiquitous RNP components to drive sex-biased immunity.

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Depletion and reconstitution of 5S RNP complexes in early pre60S ribosomes

Project description:We purified pre60S ribosomes from yeast using Nsa1-FtpA as bait in Tandem Affinity Purification experiments. We tested the depletion of RPF2, using a Galactose-depletion system, to obtain ribosomal particles depleted of 5S RNP complexes. Sample A1: Control group (wild type strain) grown in YPGal; Sample A2: Control group (wild type strain) grown in YPGlu; Sample A3: Control group (Gal::HA-RPF2 strain) grown in YPGal; and Sample A4: Test group (Gal::HA-RPF2) shifted from YPGal to YPGlu for 6 hours to achieve 5S RNP depletion. Sample N1: Purified pre-60S Nsa1-associated with 5S RNP depleted after Sucrose gradient separation; Sample N2: Purified pre-60S Nsa1-associated with 5S RNP reconstituted after Sucrose gradient separation.

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A genome-wide screen for microdeletions reveals disruption of polarity complex genes in diverse human cancers

Project description:In a genome-wide screen of human cancer cell lines, we identified homozygous intragenic microdeletions involving genes encoding components of the apical-basal cell polarity complexes. Among these, PARD3 is disrupted in cell lines and primary tumors from squamous carcinomas and glioblastomas. Reconstituting PARD3 expression in both cell types restores tight junctions and retards contact-dependent proliferation. Searching specifically for small intragenic microdeletions using high-resolution genomic arrays may be complementary to other genomic deletion screens and resequencing efforts in identifying new tumor suppressor genes. 444 human cancer cell lines of diverse tissue origin and 6 EBV-transformed B-lymphocyte cell lines (Coriell Institute for Medical Research, Camden, NJ) as controls.

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