In vivo PIWI slicing in mouse testes deviates from rules established in vitro.
MILI
MIWI
PIWI
cleavage-site mismatch
pachytene piRNA
slicer
Journal
RNA (New York, N.Y.)
ISSN: 1469-9001
Titre abrégé: RNA
Pays: United States
ID NLM: 9509184
Informations de publication
Date de publication:
03 2023
03 2023
Historique:
received:
08
07
2022
accepted:
02
12
2022
pubmed:
9
1
2023
medline:
22
2
2023
entrez:
8
1
2023
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 36617658
pii: rna.079349.122
doi: 10.1261/rna.079349.122
pmc: PMC9945443
doi:
Substances chimiques
RNA, Small Interfering
0
DNA Transposable Elements
0
RNA-Binding Proteins
0
Piwi-Interacting RNA
0
Argonaute Proteins
0
Types de publication
Letter
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
308-316Informations de copyright
© 2023 Dowling et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.
Références
Genome Biol. 2009;10(3):R25
pubmed: 19261174
Nature. 2011 Nov 27;480(7376):264-7
pubmed: 22121019
Cell Res. 2014 Jun;24(6):680-700
pubmed: 24787618
Science. 2015 May 15;348(6236):812-817
pubmed: 25977553
Nature. 2022 Aug;608(7923):618-625
pubmed: 35772669
Cell Rep. 2021 Dec 14;37(11):110110
pubmed: 34910909
Genome Res. 2021 Nov;31(11):2058-2068
pubmed: 34667116
Cell Rep. 2015 Jul 21;12(3):418-28
pubmed: 26166577
Science. 2007 May 4;316(5825):744-7
pubmed: 17446352
Elife. 2017 Oct 31;6:
pubmed: 29087293
Mol Cell. 2012 Sep 28;47(6):970-9
pubmed: 22902560
Dev Cell. 2007 Apr;12(4):503-14
pubmed: 17395546
Genes Dev. 2015 May 15;29(10):1032-44
pubmed: 25995188
Mol Cell. 2004 Jul 23;15(2):185-97
pubmed: 15260970
Nature. 2011 Oct 23;480(7376):259-63
pubmed: 22020280
Dev Cell. 2017 Jun 19;41(6):623-637.e9
pubmed: 28633017
Mol Cell. 2008 Sep 26;31(6):785-99
pubmed: 18922463
Genome Biol. 2014;15(12):550
pubmed: 25516281
Mol Cell. 2015 Sep 3;59(5):819-30
pubmed: 26340424
Nature. 2006 Jul 13;442(7099):203-7
pubmed: 16751777
Genes Dev. 2006 Jul 1;20(13):1709-14
pubmed: 16766680
Mol Cell. 2017 Oct 19;68(2):374-387.e12
pubmed: 29033321
Nat Rev Genet. 2009 Feb;10(2):94-108
pubmed: 19148191
Genes Dev. 2002 Nov 1;16(21):2733-42
pubmed: 12414724
Nature. 2020 Feb;578(7794):311-316
pubmed: 31996847
Cell. 2007 Mar 23;128(6):1089-103
pubmed: 17346786
Mol Cell. 2013 Apr 11;50(1):67-81
pubmed: 23523368
Elife. 2018 Jan 23;7:
pubmed: 29360036
Mol Cell. 2016 Jan 7;61(1):138-52
pubmed: 26669262
Nat Rev Genet. 2013 Aug;14(8):523-34
pubmed: 23797853
Science. 2006 Jul 21;313(5785):320-4
pubmed: 16809489
Mol Cell. 2011 Sep 16;43(6):1015-22
pubmed: 21925389
Nat Rev Genet. 2013 Jul;14(7):447-59
pubmed: 23732335
Science. 2015 May 15;348(6236):817-21
pubmed: 25977554
Bioinformatics. 2013 Jan 1;29(1):15-21
pubmed: 23104886
Dev Cell. 2002 Jun;2(6):819-30
pubmed: 12062093
Nat Rev Genet. 2008 Feb;9(2):102-14
pubmed: 18197166
Science. 2007 Mar 16;315(5818):1587-90
pubmed: 17322028
Nature. 2021 Sep;597(7875):285-289
pubmed: 34471284
Mol Cell. 2018 Sep 6;71(5):775-790.e5
pubmed: 30193099
Development. 2004 Feb;131(4):839-49
pubmed: 14736746
Cell Rep. 2016 Sep 13;16(11):2819-2828
pubmed: 27626653
Science. 2004 Sep 3;305(5689):1437-41
pubmed: 15284456
Nat Methods. 2015 Feb;12(2):115-21
pubmed: 25633503
PLoS Genet. 2021 Apr 8;17(4):e1009485
pubmed: 33831001
Nat Genet. 2020 Jul;52(7):728-739
pubmed: 32601478
Nat Methods. 2017 Apr;14(4):417-419
pubmed: 28263959
Nature. 2006 Jul 13;442(7099):199-202
pubmed: 16751776