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Effect of Regnase-2/MCPIP2/ZC3H12B overexpression in human glioblastoma U-251 MG cell line.

ABSTRACT: Effect of Regnase-2/MCPIP2/ZC3H12B overexpression in human glioblastoma U-251 MG cell line.

PROVIDER: PRJNA1045288 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26949793_1.fastq.gz	Fastqsanger.gz
	SRR26949793_2.fastq.gz	Fastqsanger.gz
	SRR26949794_1.fastq.gz	Fastqsanger.gz
	SRR26949794_2.fastq.gz	Fastqsanger.gz
	SRR26949795_1.fastq.gz	Fastqsanger.gz

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Effect of Regnase-2/MCPIP2/ZC3H12B overexpression in human glioblastoma U-251 MG cell line.

Project description:Regnase-2/MCPIP2/ZC3H12B is a protein belonging to the ZC3H12 protein family. There are three other proteins of this family present in humans of which Regnase-1/MCPIP1/ZC3H12A is the founder member as well as the best-described protein. All proteins of this family contain a NYN domain with endoribonuclease activity and a CCCH-type zinc finger responsible for interaction with substrate RNA. Although the NYN domain has a high degree of similarity across all four members the in vitro RNase activity was shown only for two of them, namely Regnase-1 and Regnase-3. Interestingly the ZC3H12 family proteins have unique expression patterns with Regnase-2 being expressed almost solely in the central nervous system. What is more, Regnase-2 is downregulated in human glioblastoma, and its expression is inversely correlated with tumor grade. To decipher the role of Regnase-2 in human glioblastoma we performed RNA sequencing of U-251 MG cells overexpressing Regnase-2 and control, mock-transfected cells.

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Proteomic analysis of U251 MG glioblastoma cells in the presence and absence of α-fodrin

Project description:The cortical cytoskeleton comprises mainly of the spectrin family of proteins. Spectrin and fodrin are members of this family. They support the plasma membrane. Fodrin is expressed in most tissues apart from the erythrocytes. Fodrin has been shown to be important in a wide variety of functions such as signal transduction, cardiac and brain development, etc. Through our studies it was understood that fodrin not only serves as a cortical supporting protein, but it also interacts with γ-tubulin and inhibits its microtubule nucleation activity. To gain a comprehensive view on the diverse functions of fodrin it was imperative to look into the downstream effectors of this protein in an unbiased fashion. Hence, we decided to downregulate α-fodrin in glioblastoma cells U-251 MG, and then perform a global protein expression analysis.

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Gene expression analysis of U251 human glioblastoma cell line

Project description:The human TP53 gene is frequently mutated in tumors and cell lines. Unlike other tumor suppressors that are commonly inactivated by deletions or nonsense mutations, the majority of p53-mutations are missense point mutations that result in the expression of a full-length protein with an altered amino acid that has lost sequence specific DNA-binding. Expression of mutant p53 (mutp53) confers advantages to tumor cells and transcriptional regulation of several genes mediating the beneficial effects has been shown to play a role. However, molecular mechanisms of transcriptional regulation by mutp53 are still poorly understood. We used the glioblastoma-derived U-251 MG human cell line endogenously expressing mutp53 protein (R273H mutation) to analyze gene expression profiles on Agilent Whole Human Genome Microarray after transient and stable depletion of mutp53 expression. Gene expression data was correlated with a ChIP study on a custom tiling array to understand the contribution of endogenously expressed mutp53 to transcriptional regulation. This series of microarray experiments contains the gene expression profiles of glioblastoma-derived U-251 MG human cell lines engineered to constitutively express a p53-specific shRNA or scrambled control shRNA. To reverse the effect of mutp53 depletion, stable clones were modified by stable integration of a mutp53-R273H expression construct or empty pCDNA3 vector as a control. In addition, we performed gene expression analysis of U-251 MG cells transiently transfected with p53-specific siRNA or control siRNA (3 biological replicates each).

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