Blockade of Pachytene piRNA Biogenesis Reveals a Novel Requirement for Maintaining Post-Meiotic Germline Genome Integrity

English version České info

Piwi-interacting RNAs are a diverse class of small non-coding RNAs implicated in the silencing of transposable elements and the safeguarding of genome integrity. In mammals, male germ cells express two genetically and developmentally distinct populations of piRNAs at the pre-pachytene and pachytene stages of meiosis, respectively. Pre-pachytene piRNAs are mostly derived from retrotransposons and required for their silencing. In contrast, pachytene piRNAs originate from ∼3,000 genomic clusters, and their biogenesis and function remain enigmatic. Here, we report that conditional inactivation of the putative RNA helicase MOV10L1 in mouse spermatocytes produces a specific loss of pachytene piRNAs, significant accumulation of pachytene piRNA precursor transcripts, and unusual polar conglomeration of Piwi proteins with mitochondria. Pachytene piRNA–deficient spermatocytes progress through meiosis without derepression of LINE1 retrotransposons, but become arrested at the post-meiotic round spermatid stage with massive DNA damage. Our results demonstrate that MOV10L1 acts upstream of Piwi proteins in the primary processing of pachytene piRNAs and suggest that, distinct from pre-pachytene piRNAs, pachytene piRNAs fulfill a unique function in maintaining post-meiotic genome integrity.

Vyšlo v časopise: Blockade of Pachytene piRNA Biogenesis Reveals a Novel Requirement for Maintaining Post-Meiotic Germline Genome Integrity. PLoS Genet 8(11): e32767. doi:10.1371/journal.pgen.1003038
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003038

Souhrn

Zdroje

1. SiomiMC, SatoK, PezicD, AravinAA (2011) PIWI-interacting small RNAs: The vanguard of genome defence. Nat Rev Mol Cell Biol 12: 246–258.

2. PillaiRS, ChumaS (2012) piRNAs and their involvement in male germline development in mice. Dev Growth Differ

3. AravinAA, NaumovaNM, TulinAV, VaginVV, RozovskyYM, et al. (2001) Double-stranded RNA-mediated silencing of genomic tandem repeats and transposable elements in the D. melanogaster germline. Curr Biol 11: 1017–1027.

4. BrenneckeJ, AravinAA, StarkA, DusM, KellisM, et al. (2007) Discrete small RNA-generating loci as master regulators of transposon activity in drosophila. Cell 128: 1089–1103.

5. HaaseAD, FenoglioS, MuerdterF, GuzzardoPM, CzechB, et al. (2010) Probing the initiation and effector phases of the somatic piRNA pathway in drosophila. Genes Dev 24: 2499–2504.

6. AravinAA, SachidanandamR, Bourc'hisD, SchaeferC, PezicD, et al. (2008) A piRNA pathway primed by individual transposons is linked to de novo DNA methylation in mice. Mol Cell 31: 785–799.

7. DengW, LinH (2002) Miwi, a murine homolog of piwi, encodes a cytoplasmic protein essential for spermatogenesis. Dev Cell 2: 819–830.

8. Kuramochi-MiyagawaS, KimuraT, IjiriTW, IsobeT, AsadaN, et al. (2004) Mili, a mammalian member of piwi family gene, is essential for spermatogenesis. Development 131: 839–849.

9. Kuramochi-MiyagawaS, WatanabeT, GotohK, TotokiY, ToyodaA, et al. (2008) DNA methylation of retrotransposon genes is regulated by piwi family members MILI and MIWI2 in murine fetal testes. Genes Dev 22: 908–917.

10. AravinAA, SachidanandamR, GirardA, Fejes-TothK, HannonGJ (2007) Developmentally regulated piRNA clusters implicate MILI in transposon control. Science 316: 744–747.

11. GanH, LinX, ZhangZ, ZhangW, LiaoS, et al. (2011) piRNA profiling during specific stages of mouse spermatogenesis. RNA 17: 1191–1203.

12. WatanabeT, TakedaA, TsukiyamaT, MiseK, OkunoT, et al. (2006) Identification and characterization of two novel classes of small RNAs in the mouse germline: Retrotransposon-derived siRNAs in oocytes and germline small RNAs in testes. Genes Dev 20: 1732–1743.

13. LauNC, SetoAG, KimJ, Kuramochi-MiyagawaS, NakanoT, et al. (2006) Characterization of the piRNA complex from rat testes. Science 313: 363–367.

14. GirardA, SachidanandamR, HannonGJ, CarmellMA (2006) A germline-specific class of small RNAs binds mammalian piwi proteins. Nature 442: 199–202.

15. GrivnaST, BeyretE, WangZ, LinH (2006) A novel class of small RNAs in mouse spermatogenic cells. Genes Dev 20: 1709–1714.

16. AravinA, GaidatzisD, PfefferS, Lagos-QuintanaM, LandgrafP, et al. (2006) A novel class of small RNAs bind to MILI protein in mouse testes. Nature 442: 203–207.

17. VourekasA, ZhengQ, AlexiouP, MaragkakisM, KirinoY, et al. (2012) Mili and miwi target RNA repertoire reveals piRNA biogenesis and function of miwi in spermiogenesis. Nat Struct Mol Biol 19: 773–781.

18. ReuterM, BerningerP, ChumaS, ShahH, HosokawaM, et al. (2011) Miwi catalysis is required for piRNA amplification-independent LINE1 transposon silencing. Nature 480: 264–267.

19. De FazioS, BartonicekN, Di GiacomoM, Abreu-GoodgerC, SankarA, et al. (2011) The endonuclease activity of mili fuels piRNA amplification that silences LINE1 elements. Nature 480: 259–263.

20. SaitoK, IshizuH, KomaiM, KotaniH, KawamuraY, et al. (2010) Roles for the yb body components armitage and yb in primary piRNA biogenesis in drosophila. Genes Dev 24: 2493–2498.

21. KawaokaS, IzumiN, KatsumaS, TomariY (2011) 3′ end formation of PIWI-interacting RNAs in vitro. Mol Cell 43: 1015–1022.

22. ZhengK, XiolJ, ReuterM, EckardtS, LeuNA, et al. (2010) Mouse MOV10L1 associates with piwi proteins and is an essential component of the piwi-interacting RNA (piRNA) pathway. Proc Natl Acad Sci U S A 107: 11841–11846.

23. FrostRJ, HamraFK, RichardsonJA, QiX, Bassel-DubyR, et al. (2010) MOV10L1 is necessary for protection of spermatocytes against retrotransposons by piwi-interacting RNAs. Proc Natl Acad Sci U S A 107: 11847–11852.

24. DalmayT, HorsefieldR, BraunsteinTH, BaulcombeDC (2001) SDE3 encodes an RNA helicase required for post-transcriptional gene silencing in arabidopsis. EMBO J 20: 2069–2078.

25. TomariY, DuT, HaleyB, SchwarzDS, BennettR, et al. (2004) RISC assembly defects in the drosophila RNAi mutant armitage. Cell 116: 831–841.

26. CookHA, KoppetschBS, WuJ, TheurkaufWE (2004) The drosophila SDE3 homolog armitage is required for oskar mRNA silencing and embryonic axis specification. Cell 116: 817–829.

27. VaginVV, SigovaA, LiC, SeitzH, GvozdevV, et al. (2006) A distinct small RNA pathway silences selfish genetic elements in the germline. Science 313: 320–324.

28. MaloneCD, BrenneckeJ, DusM, StarkA, McCombieWR, et al. (2009) Specialized piRNA pathways act in germline and somatic tissues of the drosophila ovary. Cell 137: 522–535.

29. OlivieriD, SykoraMM, SachidanandamR, MechtlerK, BrenneckeJ (2010) An in vivo RNAi assay identifies major genetic and cellular requirements for primary piRNA biogenesis in drosophila. EMBO J 29: 3301–3317.

30. MeisterG, LandthalerM, PetersL, ChenPY, UrlaubH, et al. (2005) Identification of novel argonaute-associated proteins. Curr Biol 15: 2149–2155.

31. ChendrimadaTP, FinnKJ, JiX, BaillatD, GregoryRI, et al. (2007) MicroRNA silencing through RISC recruitment of eIF6. Nature 447: 823–828.

32. WangPJ, McCarreyJR, YangF, PageDC (2001) An abundance of X-linked genes expressed in spermatogonia. Nat Genet 27: 422–426.

33. WangPJ, PageDC, McCarreyJR (2005) Differential expression of sex-linked and autosomal germ-cell-specific genes during spermatogenesis in the mouse. Hum Mol Genet 14: 2911–2918.

34. CarmellMA, GirardA, van de KantHJ, Bourc'hisD, BestorTH, et al. (2007) MIWI2 is essential for spermatogenesis and repression of transposons in the mouse male germline. Dev Cell 12: 503–514.

35. TanakaSS, ToyookaY, AkasuR, Katoh-FukuiY, NakaharaY, et al. (2000) The mouse homolog of drosophila vasa is required for the development of male germ cells. Genes Dev 14: 841–853.

36. SoperSF, van der HeijdenGW, HardimanTC, GoodheartM, MartinSL, et al. (2008) Mouse maelstrom, a component of nuage, is essential for spermatogenesis and transposon repression in meiosis. Dev Cell 15: 285–297.

37. MaL, BucholdGM, GreenbaumMP, RoyA, BurnsKH, et al. (2009) GASZ is essential for male meiosis and suppression of retrotransposon expression in the male germline. PLoS Genet 5: e1000635 doi:10.1371/journal.pgen.1000635.

38. ShojiM, TanakaT, HosokawaM, ReuterM, StarkA, et al. (2009) The TDRD9-MIWI2 complex is essential for piRNA-mediated retrotransposon silencing in the mouse male germline. Dev Cell 17: 775–787.

39. WatanabeT, ChumaS, YamamotoY, Kuramochi-MiyagawaS, TotokiY, et al. (2011) MITOPLD is a mitochondrial protein essential for nuage formation and piRNA biogenesis in the mouse germline. Dev Cell 20: 364–375.

40. HuangH, GaoQ, PengX, ChoiSY, SarmaK, et al. (2011) piRNA-associated germline nuage formation and spermatogenesis require MitoPLD profusogenic mitochondrial-surface lipid signaling. Dev Cell 20: 376–387.

41. SchonhoffSE, Giel-MoloneyM, LeiterAB (2004) Neurogenin 3-expressing progenitor cells in the gastrointestinal tract differentiate into both endocrine and non-endocrine cell types. Dev Biol 270: 443–454.

42. InselmanAL, NakamuraN, BrownPR, WillisWD, GouldingEH, et al. (2010) Heat shock protein 2 promoter drives cre expression in spermatocytes of transgenic mice. Genesis 48: 114–120.

43. O'GormanS, DagenaisNA, QianM, MarchukY (1997) Protamine-cre recombinase transgenes efficiently recombine target sequences in the male germ line of mice, but not in embryonic stem cells. Proc Natl Acad Sci U S A 94: 14602–14607.

44. ChumaS, HosokawaM, KitamuraK, KasaiS, FujiokaM, et al. (2006) Tdrd1/Mtr-1, a tudor-related gene, is essential for male germ-cell differentiation and nuage/germinal granule formation in mice. Proc Natl Acad Sci U S A 103: 15894–15899.

45. MeikarO, Da RosM, KorhonenH, KotajaN (2011) Chromatoid body and small RNAs in male germ cells. Reproduction 142: 195–209.

46. YabutaY, OhtaH, AbeT, KurimotoK, ChumaS, et al. (2011) TDRD5 is required for retrotransposon silencing, chromatoid body assembly, and spermiogenesis in mice. J Cell Biol 192: 781–795.

47. VasilevaA, TiedauD, FiroozniaA, Muller-ReichertT, JessbergerR (2009) Tdrd6 is required for spermiogenesis, chromatoid body architecture, and regulation of miRNA expression. Curr Biol 19: 630–639.

48. LeducF, MaquennehanV, NkomaGB, BoissonneaultG (2008) DNA damage response during chromatin remodeling in elongating spermatids of mice. Biol Reprod 78: 324–332.

49. PanJ, GoodheartM, ChumaS, NakatsujiN, PageDC, et al. (2005) RNF17, a component of the mammalian germ cell nuage, is essential for spermiogenesis. Development 132: 4029–4039.

50. RoS, ParkC, SongR, NguyenD, JinJ, et al. (2007) Cloning and expression profiling of testis-expressed piRNA-like RNAs. RNA 13: 1693–1702.

51. ChumaS, HosokawaM, TanakaT, NakatsujiN (2009) Ultrastructural characterization of spermatogenesis and its evolutionary conservation in the germline: Germinal granules in mammals. Mol Cell Endocrinol 306: 17–23.

52. ZhengK, WuX, KaestnerKH, WangPJ (2009) The pluripotency factor LIN28 marks undifferentiated spermatogonia in mouse. BMC Dev Biol 9: 38.