#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Genome-Wide and Cell-Specific Epigenetic Analysis Challenges the Role of Polycomb in Spermatogenesis


The Drosophila spermatogenesis cell differentiation pathway involves the activation of a large set of genes in primary spermatocytes. Most of these genes are activated by testis-specific TATA-binding protein associated factors (tTAFs). In the current model for the activation mechanism, Polycomb plays a key role silencing these genes in the germline precursors, and tTAF-dependent activation in primary spermatocytes involves the displacement of Polycomb from gene promoters. We investigated the genome-wide binding of Polycomb in wild type and tTAF mutant testes. According to the model we expected to see a clear enhancement in Polycomb binding at tTAF-dependent spermatogenesis genes in tTAF mutant testes. However, we find little evidence for such an enhancement in tTAF mutant testes compared to wild type. To avoid problems arising from cellular heterogeneity in whole testis analysis, we further tested the model by analysing Polycomb binding in purified germline precursors, representing cells before tTAF-dependent gene activation. Although we find Polycomb associated with its canonical targets, we find little or no evidence of Polycomb at spermatogenesis genes. The lack of Polycomb at tTAF-dependent spermatogenesis genes in precursor cells argues against a model where Polycomb displacement is the mechanism of spermatogenesis gene activation.


Vyšlo v časopise: Genome-Wide and Cell-Specific Epigenetic Analysis Challenges the Role of Polycomb in Spermatogenesis. PLoS Genet 9(10): e32767. doi:10.1371/journal.pgen.1003842
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003842

Souhrn

The Drosophila spermatogenesis cell differentiation pathway involves the activation of a large set of genes in primary spermatocytes. Most of these genes are activated by testis-specific TATA-binding protein associated factors (tTAFs). In the current model for the activation mechanism, Polycomb plays a key role silencing these genes in the germline precursors, and tTAF-dependent activation in primary spermatocytes involves the displacement of Polycomb from gene promoters. We investigated the genome-wide binding of Polycomb in wild type and tTAF mutant testes. According to the model we expected to see a clear enhancement in Polycomb binding at tTAF-dependent spermatogenesis genes in tTAF mutant testes. However, we find little evidence for such an enhancement in tTAF mutant testes compared to wild type. To avoid problems arising from cellular heterogeneity in whole testis analysis, we further tested the model by analysing Polycomb binding in purified germline precursors, representing cells before tTAF-dependent gene activation. Although we find Polycomb associated with its canonical targets, we find little or no evidence of Polycomb at spermatogenesis genes. The lack of Polycomb at tTAF-dependent spermatogenesis genes in precursor cells argues against a model where Polycomb displacement is the mechanism of spermatogenesis gene activation.


Zdroje

1. White-CooperH (2010) Molecular mechanisms of gene regulation during Drosophila spermatogenesis. Reproduction 139: 11–21 doi:10.1530/REP-09-0083

2. HillerMA, LinTY, WoodC, FullerMT (2001) Developmental regulation of transcription by a tissue-specific TAF homolog. Genes Dev 15: 1021–1030 doi:10.1101/gad.869101

3. HillerM, ChenX, PringleMJ, SuchorolskiM, SancakY, et al. (2004) Testis-specific TAF homologs collaborate to control a tissue-specific transcription program. Development 131: 5297–5308 doi:10.1242/dev.01314

4. White-CooperH, LeroyD, MacQueenA, FullerMT (2000) Transcription of meiotic cell cycle and terminal differentiation genes depends on a conserved chromatin associated protein, whose nuclear localisation is regulated. Development 127: 5463–5473.

5. JiangJ, White-CooperH (2003) Transcriptional activation in Drosophila spermatogenesis involves the mutually dependent function of aly and a novel meiotic arrest gene cookie monster. Development 130: 563–573.

6. PerezgasgaL, JiangJ, BolivalB, HillerM, BensonE, et al. (2004) Regulation of transcription of meiotic cell cycle and terminal differentiation genes by the testis-specific Zn-finger protein matotopetli. Development 131: 1691–1702 doi:10.1242/dev.01032

7. JiangJ, BensonE, BausekN, DoggettK, White-CooperH (2007) Tombola, a tesmin/TSO1-family protein, regulates transcriptional activation in the Drosophila male germline and physically interacts with always early. Development 134: 1549–1559 doi:10.1242/dev.000521

8. BeallEL, LewisPW, BellM, RochaM, JonesDL, et al. (2007) Discovery of tMAC: a Drosophila testis-specific meiotic arrest complex paralogous to Myb-Muv B. Genes Dev 21: 904–919 doi:10.1101/gad.1516607

9. DoggettK, JiangJ, AletiG, White-CooperH (2011) Wake-up-call, a lin-52 paralogue, and Always early, a lin-9 homologue physically interact, but have opposing functions in regulating testis-specific gene expression. Dev Biol 355: 381–393 doi:10.1016/j.ydbio.2011.04.030

10. KorenjakM, Taylor-HardingB, BinneUK, SatterleeJS, StevauxO, et al. (2004) Native E2F/RBF complexes contain Myb-interacting proteins and repress transcription of developmentally controlled E2F target genes. Cell 119: 181–193 doi:10.1016/j.cell.2004.09.034

11. LewisPW, BeallEL, FleischerTC, GeorletteD, LinkAJ, et al. (2004) Identification of a Drosophila Myb-E2F2/RBF transcriptional repressor complex. Genes Dev 18: 2929–2940 doi:10.1101/gad.1255204

12. GeorletteD, AhnS, MacAlpineDM, CheungE, LewisPW, et al. (2007) Genomic profiling and expression studies reveal both positive and negative activities for the Drosophila Myb MuvB/dREAM complex in proliferating cells. Genes Dev 21: 2880–2896 doi:10.1101/gad.1600107

13. SchmitF, KorenjakM, MannefeldM, SchmittK, FrankeC, et al. (2007) LINC, a human complex that is related to pRB-containing complexes in invertebrates regulates the expression of G2/M genes. Cell Cycle 6: 1903–1913.

14. AyyarS, JiangJ, ColluA, White-CooperH, WhiteRA (2003) Drosophila TGIF is essential for developmentally regulated transcription in spermatogenesis. Development 130: 2841–2852.

15. WangZ, MannRS (2003) Requirement for two nearly identical TGIF-related homeobox genes in Drosophila spermatogenesis. Development 130: 2853–2865.

16. ChenX, HillerM, SancakY, FullerMT (2005) Tissue-specific TAFs counteract Polycomb to turn on terminal differentiation. Science 310: 869–872 doi:10.1126/science.1118101

17. LanzuoloC, OrlandoV (2012) Memories from the polycomb group proteins. Annu Rev Genet 46: 561–589 doi:10.1146/annurev-genet-110711-155603

18. ChenX, LuC, PradoJR, EunSH, FullerMT (2011) Sequential changes at differentiation gene promoters as they become active in a stem cell lineage. Development 138: 2441–2450 doi:10.1242/dev.056572

19. KwongC, AdryanB, BellI, MeadowsL, RussellS, et al. (2008) Stability and dynamics of polycomb target sites in Drosophila development. PLoS Genet 4: e1000178 doi:10.1371/journal.pgen.1000178

20. LinTY, ViswanathanS, WoodC, WilsonPG, WolfN, et al. (1996) Coordinate developmental control of the meiotic cell cycle and spermatid differentiation in Drosophila males. Development 122: 1331–1341.

21. McKearinDM, SpradlingAC (1990) bag-of-marbles: a Drosophila gene required to initiate both male and female gametogenesis. Genes Dev 4: 2242–2251.

22. GonczyP, MatunisE, DiNardoS (1997) bag-of-marbles and benign gonial cell neoplasm act in the germline to restrict proliferation during Drosophila spermatogenesis. Development 124: 4361–4371.

23. GanQ, ChepelevI, WeiG, TarayrahL, CuiK, et al. (2010) Dynamic regulation of alternative splicing and chromatin structure in Drosophila gonads revealed by RNA-seq. Cell Res 20: 763–783 doi:10.1038/cr.2010.64

24. ShevelyovYY, LavrovSA, MikhaylovaLM, NurminskyID, KulathinalRJ, et al. (2009) The B-type lamin is required for somatic repression of testis-specific gene clusters. Proc Natl Acad Sci U S A 106: 3282–3287 doi:10.1073/pnas.0811933106

25. FilionGJ, van BemmelJG, BraunschweigU, TalhoutW, KindJ, et al. (2010) Systematic protein location mapping reveals five principal chromatin types in Drosophila cells. Cell 143: 212–224 doi:10.1016/j.cell.2010.09.009

26. DietzelS, NiemannH, BrucknerB, MaurangeC, ParoR (1999) The nuclear distribution of Polycomb during Drosophila melanogaster development shown with a GFP fusion protein. Chromosoma 108: 83–94.

27. MetcalfCE, WassarmanDA (2007) Nucleolar colocalization of TAF1 and testis-specific TAFs during Drosophila spermatogenesis. Dev Dyn 236: 2836–2843 doi:10.1002/dvdy.21294

28. MeiklejohnCD, LandeenEL, CookJM, KinganSB, PresgravesDC (2011) Sex Chromosome-Specific Regulation in the Drosophila Male Germline But Little Evidence for Chromosomal Dosage Compensation or Meiotic Inactivation. PLoS Biol 9: e1001126 doi:10.1371/journal.pbio.1001126

29. SurfaceLE, ThorntonSR, BoyerLA (2010) Polycomb group proteins set the stage for early lineage commitment. Cell Stem Cell 7: 288–298 doi:10.1016/j.stem.2010.08.004

30. BernsteinBE, MikkelsenTS, XieX, KamalM, HuebertDJ, et al. (2006) A bivalent chromatin structure marks key developmental genes in embryonic stem cells. Cell 125: 315–326 doi:10.1016/j.cell.2006.02.041

31. MikkelsenTS, KuM, JaffeDB, IssacB, LiebermanE, et al. (2007) Genome-wide maps of chromatin state in pluripotent and lineage-committed cells. Nature 448: 553–560 doi:10.1038/nature06008

32. StockJK, GiadrossiS, CasanovaM, BrookesE, VidalM, et al. (2007) Ring1-mediated ubiquitination of H2A restrains poised RNA polymerase II at bivalent genes in mouse ES cells. Nat Cell Biol 9: 1428–1435 doi:10.1038/ncb1663

33. BrookesE, de SantiagoI, HebenstreitD, MorrisKJ, CarrollT, et al. (2012) Polycomb associates genome-wide with a specific RNA polymerase II variant, and regulates metabolic genes in ESCs. Cell Stem Cell 10: 157–170 doi:10.1016/j.stem.2011.12.017

34. BrackenAP, DietrichN, PasiniD, HansenKH, HelinK (2006) Genome-wide mapping of Polycomb target genes unravels their roles in cell fate transitions. Genes Dev 20: 1123–1136 doi:10.1101/gad.381706

35. Román-TruferoM, Méndez-GómezHR, PérezC, HijikataA, FujimuraY, et al. (2009) Maintenance of undifferentiated state and self-renewal of embryonic neural stem cells by Polycomb protein Ring1B. Stem Cells 27: 1559–1570 doi:10.1002/stem.82

36. CarettiG, Di PadovaM, MicalesB, LyonsGE, SartorelliV (2004) The Polycomb Ezh2 methyltransferase regulates muscle gene expression and skeletal muscle differentiation. Genes Dev 18: 2627–2638 doi:10.1101/gad.1241904

37. OguroH, YuanJ, IchikawaH, IkawaT, YamazakiS, et al. (2010) Poised lineage specification in multipotential hematopoietic stem and progenitor cells by the polycomb protein Bmi1. Cell Stem Cell 6: 279–286 doi:10.1016/j.stem.2010.01.005

38. BoyerLA, PlathK, ZeitlingerJ, BrambrinkT, MedeirosLA, et al. (2006) Polycomb complexes repress developmental regulators in murine embryonic stem cells. Nature 441: 349–353 doi:10.1038/nature04733

39. LeeTI, JennerRG, BoyerLA, GuentherMG, LevineSS, et al. (2006) Control of developmental regulators by Polycomb in human embryonic stem cells. Cell 125: 301–313 doi:10.1016/j.cell.2006.02.043

40. SchwartzYB, KahnTG, NixDA, LiXY, BourgonR, et al. (2006) Genome-wide analysis of Polycomb targets in Drosophila melanogaster. Nat Genet 38: 700–705 doi:10.1038/ng1817

41. TolhuisB, de WitE, MuijrersI, TeunissenH, TalhoutW, et al. (2006) Genome-wide profiling of PRC1 and PRC2 Polycomb chromatin binding in Drosophila melanogaster. Nat Genet 38: 694–699 doi:10.1038/ng1792

42. LawrencePA, JohnstonP, StruhlG (1983) Different requirements for homeotic genes in the soma and germ line of Drosophila. Cell 35: 27–34.

43. GanQ, SchonesDE, Ho EunS, WeiG, CuiK, et al. (2010) Monovalent and unpoised status of most genes in undifferentiated cell-enriched Drosophila testis. Genome Biol 11: R42 doi:10.1186/gb-2010-11-4-r42

44. SanoH, NakamuraA, KobayashiS (2002) Identification of a transcriptional regulatory region for germline-specific expression of vasa gene in Drosophila melanogaster. Mech Dev 112: 129–139.

45. McKearinD, OhlsteinB (1995) A role for the Drosophila bag-of-marbles protein in the differentiation of cystoblasts from germline stem cells. Development 121: 2937–2947.

46. DahlJA, CollasP (2008) A rapid micro chromatin immunoprecipitation assay (microChIP). Nat Protoc 3: 1032–1045 doi:10.1038/nprot.2008.68

47. DahlJA, CollasP (2008) MicroChIP–a rapid micro chromatin immunoprecipitation assay for small cell samples and biopsies. Nucleic Acids Res 36: e15 doi:10.1093/nar/gkm1158

48. PappB, MullerJ (2006) Histone trimethylation and the maintenance of transcriptional ON and OFF states by trxG and PcG proteins. Genes Dev 20: 2041–2054 doi:10.1101/gad.388706

49. BentonR, PalaciosIM, St JohnstonD (2002) Drosophila 14-3-3/PAR-5 is an essential mediator of PAR-1 function in axis formation. Dev Cell 3: 659–671.

50. ChintapalliVR, WangJ, DowJAT (2007) Using FlyAtlas to identify better Drosophila melanogaster models of human disease. Nat Genet 39: 715–720 doi:10.1038/ng2049

51. RedhouseJL, MozziconacciJ, WhiteRA (2011) Co-transcriptional architecture in a Y loop in Drosophila melanogaster. Chromosoma 120: 399–407 doi:10.1007/s00412-011-0321-1

52. KosmanD, MizutaniCM, LemonsD, CoxWG, McGinnisW, et al. (2004) Multiplex detection of RNA expression in Drosophila embryos. Science 305: 846 doi:10.1126/science.1099247

53. LanglaisKK, BrownJL, KassisJA (2012) Polycomb Group Proteins Bind an engrailed PRE in Both the “ON” and “OFF” Transcriptional States of engrailed. PLoS ONE 7: e48765 doi:10.1371/journal.pone.0048765

Štítky
Genetika Reprodukčná medicína

Článok vyšiel v časopise

PLOS Genetics


2013 Číslo 10
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#