In vivo PIWI slicing in mouse testes deviates from rules established in vitro

Article Article scientifique Lettre In vivo PIWI slicing in mouse testes deviates from rules established in vitro eng 2022-12-13 2023-03-13 author Dowling Mark 0009-0006-2091-0724 877696 Département de biologie moléculaire et cellulaire (MOCEL) author Homolka David 856314 Département de biologie moléculaire et cellulaire (MOCEL) author Raad Nicole 989097 Département de biologie moléculaire et cellulaire (MOCEL) author Gos Pascal 27819 Département de biologie moléculaire et cellulaire (MOCEL) author Pandey Radha Raman 0000-0002-4439-8386 853296 author Pillai Ramesh 854560 Département de biologie moléculaire et cellulaire (MOCEL) 308-316 10.1261/rna.079349.122 36617658 1355-8382 PMC9945443 http://rnajournal.cshlp.org/lookup/doi/10.1261/rna.079349.122 <p>Argonautes are small RNA-binding proteins, with some having small RNA-guided endonuclease (slicer) activity that cleaves target nucleic acids. One cardinal rule that is structurally defined is the inability of slicers to cleave target RNAs when nucleotide mismatches exist between the paired small RNA and the target at the cleavage site. Animal-specific PIWI clade Argonautes associate with PIWI-interacting RNAs (piRNAs) to silence transposable elements in the gonads, and this is essential for fertility. We previously demonstrated that purified endogenous mouse MIWI fails to cleave mismatched targets in vitro. Surprisingly, here we find using knock-in mouse models that target sites with cleavage-site mismatches at the 10th and 11th piRNA nucleotides are precisely sliced in vivo. This is identical to the slicing outcome in knock-in mice where targets are base-paired perfectly with the piRNA. Additionally, we find that pachytene piRNA-guided slicing in both these situations failed to initiate phased piRNA production from the specific target mRNA we studied. Instead, the two slicer cleavage fragments were retained in PIWI proteins as pre-piRNA and 17–19 nt by-product fragments. Our results indicate that PIWI slicing rules established in vitro are not respected in vivo, and that all targets of PIWI slicing are not substrates for piRNA biogenesis.</p> MILI MIWI PIWI Cleavage-site mismatch Pachytene piRNA Slicer Dewey 570 Republic and Canton of Geneva Swiss National Science Foundation Gene regulation by RNA modification writers and readers 207468 CH Projects Swiss National Science Foundation GermMethylation (ERC-2014-CoG) 166923 CH Programmes 43 Département de biologie moléculaire 161 false RNA 29 3 cec8b434-a5c3-4cc9-8753-7a6f962d7aaa Article (Published version) Public Article-scientifique-from-unpaywall.pdf 955203 application/pdf CC BY-4.0 5d4daf1a8654d5ef1e68e3ff76a123db727219051f5f4b9db1e248602ee3d4dd openAccess Faculté des sciences / Section de biologie / Département de biologie moléculaire

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In vivo PIWI slicing in mouse testes deviates from rules established in vitro	RNA			2023	93	235

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Raad, Nicole