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Roles of the kinase NUAK1 on spliceosome in the mouse cortex

ABSTRACT: Roles of the kinase NUAK1 on spliceosome in the mouse cortex

PROVIDER: PRJNA941251 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR23707000_1.fastq.gz	Fastqsanger.gz
	SRR23707000_2.fastq.gz	Fastqsanger.gz
	SRR23707001_1.fastq.gz	Fastqsanger.gz
	SRR23707001_2.fastq.gz	Fastqsanger.gz
	SRR23707002_1.fastq.gz	Fastqsanger.gz

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Roles of the kinase NUAK1 on spliceosome in the mouse cortex

Project description:The project focuses on the protein kinase NUAK1, which controls the development of brain cortical connectivity in mice. Recent data suggest that NUAK1 is involved in the regulation of the spliceosome and more broadly of mRNA biology. The aim of this project is to test whether inactivation of NUAK1 (constitutive KO) affects the expression level of genes relevant for cortical development in the embryonic brain. We collected the cortex of embryos (littermates) at the late embryonic developmental stage (E18.5, i.e. just before birth). The samples will be either KO or WT (controls). We collected the samples on the same day, under the same conditions, and as far as possible we made homogeneous groups: same litter, same proportion of males/females.

2024-01-22 | GSE226698 | GEO

Local inhibition of protein phosphatase 1 by NUAK1 promotes spliceosome assembly and reveals a MYC-sensitive feedback control of transcriptional elongation

Project description:Deregulated expression of MYC induces a dependence on the NUAK1 kinase, but the molecular mechanisms underlying this dependence have not been fully clarified. Here we show that NUAK1 is a predominantly nuclear protein that associates with a network of nuclear protein phosphatase 1 (PP1) interactors and that PNUTS, a nuclear regulatory subunit of PP1, is phosphorylated by NUAK1. Both NUAK1 and PNUTS associate with the splicing machinery. Inhibition of NUAK1 abolishes chromatin association of PNUTS, reduces spliceosome activity and suppresses nascent RNA synthesis. Activation of MYC does not bypass the requirement for NUAK1 for spliceosome activity, but significantly attenuates transcription inhibition. Consequently, NUAK1 inhibition in MYC-transformed cells induces global accumulation of RNAPII both at the pause-site and at the first exon/intron boundary, but does not increase mRNA synthesis. We suggest that NUAK1 inhibition in the presence of deregulated MYC traps non-productive RNAPII due to the absence of correctly assembled spliceosomes.

2020-02-01 | GSE129925 | GEO

Transcriptomic profiling in NUAK1 KO neurons

Project description:The project focuses on the protein kinase NUAK1, which controls the development of brain cortical connectivity in mice. Recent data suggest that NUAK1 is involved in the regulation of the spliceosome and more broadly of mRNA biology. The aim of this project is to test whether inactivation of NUAK1 (constitutive KO) in primary neuronal cultures affects the expression level of genes relevant for cortical and axon development. We prepared primary cultures of mouse neurons at the embryonic developmental stage E15.5. The samples were either KO or WT (controls). We collected the samples from 4 individual cultures performed blind to genotype, then selected the samples based on PCR to identify sex and genotype. Cultures were thus performed under the same conditions but not performed the same day nor from the same litter for each sample.

2024-01-22 | GSE227203 | GEO

Colorectal tumors require NUAK1 for protection from oxidative stress

Project description:NUAK1 is a member of the AMPK family of kinases involved in regulation of cell adhesion, metabolism and stress responses. Our previous analysis revealed a requirement for NUAK1 to sustain viability when MYC is overexpressed. Human colorectal cancer (CRC) shows near uniform overexpression of MYC, suggesting that this cancer may be particularly amenable to NUAK1 inhibition. This work reveals that elevated NUAK1 expression in human CRC correlates with more aggressive disease, lymph node metastasis and reduced overall survival. Deletion of Nuak1 in murine intestines prevented outgrowth of colonic tumors driven by loss of Apc and KRas mutation, while depletion of Nuak1 in pre-existing tumors drove rapid tumor regression. Mechanistically, we show that NUAK1 promotes anti-oxidant gene expression by facilitating nuclear import of NRF2. Deletion or inhibition of NUAK1 renders cells vulnerable to oxidative stress and provision of exogenous anti-oxidants protects cells in culture and colonic tumors in situ from Nuak1 depletion.

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In situ depletion of Nuak1 in murine colon tumours

Project description:The aim of the project is to determine the impact of NUAK1 depletion on the transcriptome of in situ colon tumours. Tumours were initiated in Villin-CRE-ER;APC-floxed;DI-shNUAK1 mice, followed by treatment for 1 week with dextran sodium sulphate. 68 days after tumour initiation, NUAK1 shRNAs were induced by doxycycline administration (via gavage), or left un-induced. 6 tumours were harvested from NUAK1 shRNA expressing mice and 6 from controls. RNA was isolated using Qiagen RNEasy and analyzed by Illumina paired-end RNA-SEQ

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Project description:Spliceosome iCLIP

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Project description:The intent of the experiment is to determine how much of NRF2 (NFE2L2)-dependent transcription requires NUAK1 upstream. SW480 cells were transfected with siRNA targeting either NRF2, NUAK1 or non-targeting control, and harvested for analysis by RNA-SEQ 48hrs after transfection. mRNA was analysed by Illumina paired-end RNA-SEQ

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2019-08-30 | E-MTAB-6950 | biostudies-arrayexpress

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