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Effect of depletion of A3B on gene expression of 92.1 cells and overexpression of A3B on gene expression of MEL290 cells

ABSTRACT: Effect of depletion of A3B on gene expression of 92.1 cells and overexpression of A3B on gene expression of MEL290 cells

PROVIDER: PRJNA905401 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR22408342_1.fastq.gz	Fastqsanger.gz
	SRR22408342_2.fastq.gz	Fastqsanger.gz
	SRR22408343_1.fastq.gz	Fastqsanger.gz
	SRR22408343_2.fastq.gz	Fastqsanger.gz
	SRR22408344_1.fastq.gz	Fastqsanger.gz

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Effect of depletion of A3B on gene expression of 92.1 cells and overexpression of A3B on gene expression of MEL290 cells

Project description:To investigate the function of A3B, we established 92.1 cell lines with A3B knocked down and MEL290 cell lines with A3B overexpression. We then performed gene expression profiling analysis using data obtained from RNA-seq of 4 different cell lines with two replicates.

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Decoding the Genes that A3B regulated

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Project description:Mutations in cancer are due in part to DNA cytosine deamination by APOBEC3B (A3B). While low in healthy tissues, A3B expression and activity are elevated in tumors and further increase in metastases. However, molecular mechanisms responsible for A3B transcriptional regulation are poorly understood. Here, we address whether the RB/E2F pathway, which is often dysregulated in breast cancer, is a molecular trigger of A3B overexpression. First, an A3B promoter-driven luciferase reporter was used to demonstrate reporter activation by disruption of only one out of five predicted E2F binding sites. Second, A3B-luciferase reporter activation was also triggered by expressing the BK polyomavirus T-antigen, which inactivates RB and thereby alleviates repression of E2F regulated genes. Importantly, A3B-luciferase reporter induction by BK polyomavirus T-antigen could not be further increased by mutating the functional E2F binding site. Third, both CRISPR disruption and targeted base substitutions in the endogenous E2F binding site caused strong A3B upregulation and confirmed importance of this regulatory element. Fourth, proteomics experiments showed that members of the DREAM and PRC1.6-complexes including multiple E2F family members are able to bind to wild-type but not E2F mutant promoter sequences. Finally, a combination of genetic and biochemical studies implicated E2F4 and E2F6 in endogenous A3B gene repression. Altogether, our studies demonstrate that A3B expression is suppressed in normal cells by the RB/E2F axis and that viral or mutational disruption of this pathway causes overexpression of this DNA deaminase in cancer and contributes mutational fuel to tumor evolution.

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Effect of overexpression of A3B and RNA editing calling in mouse tissues

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