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Saccharomyces cerevisiae BY4741

ABSTRACT: Genome-wide base editor screen identifies regulators of protein abundance in yeast

PROVIDER: PRJNA732764 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14727004_1.fastq.gz	Fastqsanger.gz
	SRR14727004_2.fastq.gz	Fastqsanger.gz
	SRR14727005_1.fastq.gz	Fastqsanger.gz
	SRR14727005_2.fastq.gz	Fastqsanger.gz
	SRR14727006_1.fastq.gz	Fastqsanger.gz

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Genome-wide base editor screen identifies regulators of protein abundance in yeast

Project description:Proteins are key molecular players in a cell, and their abundance is extensively regulated not just at the level of gene expression but also post-transcriptionally. Here, we describe a genetic screen in yeast that enables systematic characterization of how protein abundance regulation is encoded in the genome. The screen combines a CRISPR/Cas9 base editor to introduce point mutations with fluorescent tagging of endogenous proteins to facilitate a flow-cytometric readout. We first benchmarked base editor performance in yeast with individual gRNAs as well as in positive and negative selection screens. We then examined the effects of 16,452 genetic perturbations on the abundance of eleven proteins representing a variety of cellular functions. We uncovered hundreds of regulatory relationships, including a novel link between the GAPDH isoenzymes Tdh1/2/3 and the Ras/PKA pathway. Many of the identified regulators are specific to one of the eleven proteins, but we also found genes that, upon perturbation, affected the abundance of most of the tested proteins. While the more specific regulators usually act transcriptionally, broad regulators often have roles in protein translation. Overall, our novel screening approach provides unprecedented insights into the components, scale and connectedness of the protein regulatory network.

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