A mating-type mutagenesis screen identifies a zinc-finger protein required for specific DNA excision events in Paramecium.

Nucleic Acids Research
Simran BhullarEric Meyer

Abstract

In the ciliate Paramecium tetraurelia, functional genes are reconstituted during development of the somatic macronucleus through the precise excision of ∼45 000 single-copy Internal Eliminated Sequences (IESs), thought to be the degenerate remnants of ancient transposon insertions. Like introns, IESs are marked only by a weak consensus at their ends. How such a diverse set of sequences is faithfully recognized and precisely excised remains unclear: specialized small RNAs have been implicated, but in their absence up to ∼60% of IESs are still correctly excised. To get further insight, we designed a mutagenesis screen based on the hypersensitivity of a specific excision event in the mtA gene, which determines mating types. Unlike most IES-containing genes, the active form of mtA is the unexcised one, allowing the recovery of hypomorphic alleles of essential IES recognition/excision factors. Such is the case of one mutation recovered in the Piwi gene PTIWI09, a key player in small RNA-mediated IES recognition. Another mutation identified a novel protein with a C2H2 zinc finger, mtGa, which is required for excision of a small subset of IESs characterized by enrichment in a 5-bp motif. The unexpected implication of a sequence-specif...Continue Reading

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Citations

May 29, 2020·Cellular and Molecular Life Sciences : CMLS·Iwona RzeszutekMariusz Nowacki
Jan 26, 2022·The Journal of Eukaryotic Microbiology·Eric Cole, Jacek Gaertig

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Datasets Mentioned

BETA
PRJEB27011

Methods Mentioned

BETA
genotyping
PCR

Software Mentioned

bowtie2
DREME
clustalW
MUSCLE
cutadapt

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