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Unbiased screen of RNA tailing enzymes reveals a poly(UG) polymerase required for genome integrity and RNA silencing

ABSTRACT: Unbiased screen of RNA tailing enzymes reveals a poly(UG) polymerase required for genome integrity and RNA silencing

PROVIDER: PRJNA508883 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8290490_1.fastq.gz	Fastqsanger.gz
	SRR8290490_2.fastq.gz	Fastqsanger.gz
	SRR8290500_1.fastq.gz	Fastqsanger.gz
	SRR8290500_2.fastq.gz	Fastqsanger.gz
	SRR8290510_1.fastq.gz	Fastqsanger.gz

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Unbiased screen of RNA tailing enzymes reveals a poly(UG) polymerase required for genome integrity and RNA silencing

Project description:Ribonucleotidyl transferases (rNTases) add non-templated ribonucleotides to diverse RNAs. We developed a screening strategy in S. cerevisiae to identify sequences added by candidate enzymes from different organisms at single-nucleotide resolution. The rNTase activities of 19 previously unexplored enzymes were determined. In addition to poly(A)- and poly(U)-adding enzymes, we identified a C-adding enzyme that is likely part of a two-enzyme system that adds CCA to tRNAs in a eukaryote; a nucleotidyl transferase that adds nucleotides to RNA without apparent nucleotide preference; and a poly(UG) polymerase, C. elegans MUT-2, which adds alternating U and G nucleotides to form poly(UG) tails. MUT-2 is known to be required for certain forms of RNA silencing, and mutations in the enzyme that are defective in silencing also fail to add poly(UG) tails in our assay. We propose that MUT-2 poly(UG) polymerase activity is required to promote genome integrity and RNA silencing.

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Project description:Ribonucleotidyl transferases (rNTases) add untemplated ribonucleotides to diverse RNAs. We have developed TRAID-seq, a screening strategy in Saccharomyces cerevisiae to identify sequences added to a reporter RNA at single-nucleotide resolution by overexpressed candidate enzymes from different organisms. The rNTase activities of 22 previously unexplored enzymes were determined. In addition to poly(A)- and poly(U)-adding enzymes, we identified a cytidine-adding enzyme that is likely to be part of a two-enzyme system that adds CCA to tRNAs in a eukaryote; a nucleotidyl transferase that adds nucleotides to RNA without apparent nucleotide preference; and a poly(UG) polymerase, Caenorhabditis elegans MUT-2, that adds alternating uridine and guanosine nucleotides to form poly(UG) tails. MUT-2 is known to be required for certain forms of RNA silencing, and mutants of the enzyme that result in defective silencing did not add poly(UG) tails in our assay. We propose that MUT-2 poly(UG) polymerase activity is required to promote genome integrity and RNA silencing.

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