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Chromosoma

, Volume 117, Issue 1, pp 25–39 | Cite as

The trithorax group and Pc group proteins are differentially involved in heterochromatin formation in Drosophila

  • Laura FantiEmail author
  • Barbara Perrini
  • Lucia Piacentini
  • Maria Berloco
  • Enzo Marchetti
  • Gioacchino Palumbo
  • Sergio PimpinelliEmail author
Research Article

Abstract

In Drosophila, the Polycomb group and trithorax group proteins play a critical role in controlling the expression states of homeotic gene complexes during development. The common view is that these two classes of proteins bind to the homeotic complexes and regulate transcription at the level of chromatin. In the present work, we tested the involvement of both groups in mitotic heterochromatin formation in Drosophila. Using specific antibodies, we show that some of the tested Pc-G proteins are present in heterochromatin, while all the tested trx-G proteins localize to specific regions of heterochromatin in both mitotic chromosomes and interphase nuclei. We also observed that mutations in trx-G genes are recessive enhancers of position-effect variegation and are able to repress the transcription of heterochromatic genes. These results strongly suggest that trx-G proteins, along with some Pc-G proteins, play an active role in heterochromatin formation in Drosophila.

Keywords

Polytene Chromosome Mitotic Chromosome Heterochromatic Region Homeotic Gene Heterochromatin Formation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We thank K. Golic G. Karpen, T. Lyttle for their comments on manuscript. We thank C. Goday for the useful suggestions in elaborating some of the immunostaining procedures. We thank G. Cavalli, J. Eissenberg, M. Gatti, R. Johns, R. Paro, G. Reuter, A. Shearn, J. Tamkun, and Bloomington Stocks Center for the mutant Drosophila strains. We thank K. Ahmad, S. Elgin, P. Harte, S. Henikoff, R. Johns, G. Karpen, J. Kassis, A. Mazo, R. Paro, V. Pirrotta, A. G. Reuter, A. Shearn, J. Tamkun, C. Wu for antibodies.

References

  1. Bantignies F, Goodman RH, Smolik SM (2000) Functional interaction between the coactivator Drosophila CREB-binding protein and ASH1, a member of the trithorax group of chromatin modifiers. Mol Cell Biol 20:9317–9330PubMedCrossRefGoogle Scholar
  2. Bejarano F, Busturia A (2004) Function of trithorax-like gene during Drosophila development. Dev Biol 268:327–341PubMedCrossRefGoogle Scholar
  3. Belsel C, Imhof A, Greene J, Kremmer E, Sauer F (2002) Histone methylation by the Drosophila epigenetic transcriptional regulator Ash1. Nature 419:857–862CrossRefGoogle Scholar
  4. Bhat KM, Farkas G, Karch F, Gyurkovics H, Gausz J, Schedl P (1996) The GAGA factor is required in the early Drosophila embryo not only for transcriptional regulation but also for nuclear division. Development 122:1113–1124PubMedGoogle Scholar
  5. Brown JL, Mucci D, Whiteley M, Dirksen M-L, Kassis JA (1998) The Drosophila Polycomb group gene pleiohomeotic encodes a sequence-specific DNA binding protein with homology to the multifunctional mammalian transcription factor YY1. Mol Cell 1:1057–1064PubMedCrossRefGoogle Scholar
  6. Buchenau P, Hodgson J, Strutt H, Arndt-Jovin DJ (1998) The distribution of polycomb-group proteins during cell division and development in Drosophila embryos: impact on models for silencing. J Cell Biol 141:469–481PubMedCrossRefGoogle Scholar
  7. Busturia A, LLoyd A, Bejarano F, Zavortink M, Xin H, Sakonju S (2001) The MCP silencer of the Drosophila Abd-B gene requires both pleiohomeotic and GAGA factor for the maintenance of repression. Development 128:2163–2173PubMedGoogle Scholar
  8. Cairns BR (1998) Chromatin remodeling machines: similar motors, ulterior motives. Trends Biochem Sci 23:20–25PubMedCrossRefGoogle Scholar
  9. Cavalli G, Paro R (1999) Epigenetic inheritance of active chromatin after removal of the main transactivator. Science 286:955–958PubMedCrossRefGoogle Scholar
  10. Chinwalla V, Jane PE, Harte PJ (1995) The Drosophila trithorax protein binds to specific chromosomal sites and is co-localized with Polycomb at many sites. EMBO J 14:2056–2065PubMedGoogle Scholar
  11. Cryderman DE, Grade SK, Li Y, Fanti L, Pimpinelli S, Wallrath LL (2005) Role of Drosophila HP1 in euchromatic gene expression. Dev Dyn 232:767–774PubMedCrossRefGoogle Scholar
  12. Czermin B, Melfi R, McCabe D, Seitz V, Imhof A, Pirrotta V (2002) Drosophila enhancer of Zeste/ESC complexes have a histone H3 methyltransferase activity that marks chromosomal Polycomb sites. Cell 111:185–196PubMedCrossRefGoogle Scholar
  13. DeCamillis M, Cheng NS, Pierre D, Brock H (1992) The polyhomeotic gene of Drosophila encodes a chromatin protein that shares polytene chromosome-binding sites with Polycomb. Genes Dev 6:223–232PubMedCrossRefGoogle Scholar
  14. Deuring R, Fanti L, Armstrong JA, Sarte M, Papoulas O, Prestel M, Daubresse G, Verardo M, Moseley SL, Berloco M, Tsukiyama T, Wu C, Pimpinelli S, Tamkun JW (2000) The ISWI chromatin-remodeling protein is required for gene expression and the maintenance of higher order chromatin structure in vivo. Mol Cell 5:355–365PubMedCrossRefGoogle Scholar
  15. Duncan IM (1982) Polycomblike: a gene that appears to be required for the normal expression of the bithorax and antennapedia gene complexes of Drosophila melanogaster. Genetics 102:49–70PubMedGoogle Scholar
  16. Dorn R, Krauss V, Reuter G, Saumweber H (1993) The enhancer of position-effect variegation of Drosophila, E(var)3-93D, codes for a chromatin protein containing a conserved domain common to several transcriptional regulators. Proc Natl Acad Sci USA 90:11376–11380PubMedCrossRefGoogle Scholar
  17. Eissenberg JC, James TC, Foster-Hartnett DM, Hartnett T, Ngan V, Elgin SCR (1990) Mutation in a heterochromatin-specific chromosomal protein is associated with suppression of position-effect variegation in Drosophila melanogaster. Proc Natl Acad Sci USA 87:9923–9927PubMedCrossRefGoogle Scholar
  18. Elfring LK, Deuring R, McCallum CM, Peterson CL, Tamkun JW (1994) Identification and characterization of Drosophila relatives of the yeast transcriptional activator SNF2/SWI2. Mol Cell Biol 14:2225–2234PubMedGoogle Scholar
  19. Fanti L, Pimpinelli S (2004) Immunostaining of squash preparations of chromosomes of larval brains. Methods Mol Biol 247:353–361PubMedGoogle Scholar
  20. Fanti L, Giovinazzo G, Berloco M, Pimpinelli S (1998) The heterochromatin protein 1 (HP1) prevents telomere fusions in Drosophila melanogaster. Mol Cell 2:1–20CrossRefGoogle Scholar
  21. Farkas G, Janos G, Galloni M, Reuter G, Gyurkovics H, Karch F (1994) The Trithorax-like gene encodes the Drosophila GAGA factor. Nature 371:806–808PubMedCrossRefGoogle Scholar
  22. Franke A, DeCamillis M, Zink D, Cheng N, Brock H, Paro R (1992) Polycomb and Polyhomeotic are constituents of a multimeric protein complex in chromatin of Drosophila melanogaster. EMBO J 11:2941–2950PubMedGoogle Scholar
  23. Fritsch C, Brown JL, Kassis JA, Müller J (1999) The DNA-binding Polycomb group protein Pleiohomeotic mediates silencing of a Drosophila homeotic gene. Development 126:3905–3913PubMedGoogle Scholar
  24. Gatti M, Pimpinelli S (1992) Functional elements in Drosophila melanogaster heterochromatin. Annu Rev Genet 26:239–275PubMedCrossRefGoogle Scholar
  25. Horard B, Tatout C, Poux S, Pirrotta V (2000) Structure of a polycomb response element and in vitro binding of polycomb group complexes containing GAGA factor. Mol Cell Biol 20:3187–3197PubMedCrossRefGoogle Scholar
  26. Ingham PW (1985) A clonal analysis of the requirement for the trithorax gene in the diversification of segments in Drosophila. J Embryol Exp Morphol 89:349–365PubMedGoogle Scholar
  27. James TC, Elgin SCR (1986) Identification of nonhistone chromosomal protein associated with heterochromatin in Drosophila and its gene. Mol Cell Biol 6:3862–3872PubMedGoogle Scholar
  28. James TC, Eissenberg JC, Craig C, Dietrich V, Hobson A, Elgin SCR (1989) Distribution patterns of HP1, a heterochromatin-associated nonhistone chromosomal protein of Drosophila. Eur J Cell Biol 50:170–180PubMedGoogle Scholar
  29. Jenuwein T (2001) Re-SET-ting heterochromatin by histone methyltransferases. Trends Cell Biol 11:266–273PubMedCrossRefGoogle Scholar
  30. Jones RS, Gelbart WM (1993) The Drosophila Polycomb-group gene Enhancer of zeste contains a region with sequence similarity to trithorax. Mol Cell Biol 13:6357–6366PubMedGoogle Scholar
  31. Jürgens G (1985) A group of genes controlling the spatial expression of the bithorax complex in Drosophila. Nature 316:153–155CrossRefGoogle Scholar
  32. Kennison JA (1993) Transcriptional activation of Drosophila homeotic genes from distant regulatory elements. Trends Genet 9:75–79PubMedCrossRefGoogle Scholar
  33. Kuzin B, Tillib S, Sedkov Y, Mizrokhi L, Mazo A (1994) The Drosophila trithorax gene encodes a chromosomal protein and directly regulates the region-specific homeotic gene fork head. Genes Dev 8:2478–2490PubMedCrossRefGoogle Scholar
  34. Laible G, Wolf A, Dorn R, Reuter G, Nislow C, Lebersorger A, Popkin D, Pillus L, Jenuwein T (1997) Mammalian homologues of the Polycomb-group gene Enhancer of zeste mediate gene silencing in Drosophila heterochromatin and at S. cerevisiae telomeres. EMBO J 16:3219–3232PubMedCrossRefGoogle Scholar
  35. LaJeunesse D, Shearn A (1996) E(z): a Polycomb group gene or a trithorax group gene? Development 122:2189–2197PubMedGoogle Scholar
  36. Lohe AR, Hilliker AJ, Roberts PA (1993) Mapping simple repeated DNA sequences in heterochromatin of Drosophila melanogaster. Genetics 134:1149–1174PubMedGoogle Scholar
  37. Lu BY, Bishop CP, Eissenberg JC (1996) Developmental timing and tissue specificity of heterochromatin-mediated silencing. EMBO J 15:1323–1332PubMedGoogle Scholar
  38. Lu BY, Emtage PC, Duyf BJ, Hilliker AJ, Eissenberg JC (2000) Heterochromatin protein 1 is required for the normal expression of two heterochromatin genes in Drosophila. Genetics 155:699–708PubMedGoogle Scholar
  39. Martin EC, Adler PN (1993) The Polycomb group gene posterior sex combs encodes a chromosomal protein. Development 117:641–655PubMedGoogle Scholar
  40. Martínez-Balbás MA, Dey A, Rabindran SK, Ozato K, Wu C (1995) Displacement of sequence-specific transcription factors from mitotic chromatin. Cell 83:29–38PubMedCrossRefGoogle Scholar
  41. Mazo AM, Huang DH, Mozer BA, Dawid IB (1990) The trithorax gene, a trans-acting regulator of the bithorax complex in Drosophila, encodes a protein with zinc-binding domains. Proc Natl Acad Sci USA 87:2112–2116PubMedCrossRefGoogle Scholar
  42. McDonald JF (1990) Macroevolution and retroviral elements. Bioscience 40:183–191CrossRefGoogle Scholar
  43. Orlando V, Paro R (1993) Mapping Polycomb-repressed domains in the bithorax complex using in vivo formaldehyde cross-linked chromatin. Cell 75:1187–1198PubMedCrossRefGoogle Scholar
  44. Paranjape SM, Kamakaka RT, Kadonaga JT (1994) Role of chromatin structure in the regulation of transcription by RNA polymerase II. Ann Rev Biochem 63:265–297PubMedCrossRefGoogle Scholar
  45. Paro R (1990) Imprinting a determinated state into the chromatin of Drosophila. Trends Genet 6:416–421PubMedCrossRefGoogle Scholar
  46. Petruk S, Sedkov Y, Smith S, Tillib S, Kraevski V, Nakamura T, Canaani E, Croce MC, Mazo A (2001) Trithorax and dCBP acting in a complex to maintain expression of a homeotic gene. Science 294:1331–1334PubMedCrossRefGoogle Scholar
  47. Petruk S, Sedkov Y, Riley KM, Hodgson J, Schweisguth F, Hirose S, Jaynes JB, Brock HW, Mazo A (2006) transcription of bxd noncoding RNAs promoted by Trithorax represses Ubx in cis by transcriptional interference. Cell 127:1209–1221PubMedCrossRefGoogle Scholar
  48. Pimpinelli S (2000) The functional and structural organization of Drosophila heterochromatin. Chromosome Today 13:29–43Google Scholar
  49. Pimpinelli S, Santini G, Gatti M (1978) 3H-Actinomycin-D binding to mitotic chromosomes of Drosophila melanogaster. Chromosoma 66:389–395PubMedCrossRefGoogle Scholar
  50. Pimpinelli S, Berloco M, Fanti L, Dimitri P, Bonaccorsi S, Marchetti E, Caizzi R, Caggese C, Gatti M (1995) Transposable elements are stable structural components of Drosophila melanogaster heterochromatin. Proc Natl Acad Sci USA 92:3804–3808PubMedCrossRefGoogle Scholar
  51. Pirrotta V (1997) Pc-G complexes and chromatin silencing. Curr Opin Genet Dev 7:249–258PubMedCrossRefGoogle Scholar
  52. Platero JS, Hartnett T, Eissenberg JC (1995) Functional analysis of the chromo domain of HP1. EMBO J 14:3977–3986PubMedGoogle Scholar
  53. Platero JS, Csink AK, Quintanilla A, Henikoff S (1998) Changes in chromosomal localization of heterochromatin binding proteins during the cell cycle in Drosophila. J Cell Biol 140:1297–1306PubMedCrossRefGoogle Scholar
  54. Raff JW, Kellum R, Alberts B (1994) The Drosophila GAGA transcription factor is associated with specific regions of heterochromatin throughout the cell cycle. EMBO J 13:5977–5983PubMedGoogle Scholar
  55. Rastelli L, Chan CS, Pirrotta V (1993) Related chromosome binding sites for zeste, suppressor of zeste and Polycomb group proteins in Drosophila and their dependence on Enhancer of zeste function. EMBO J 12:1513–1522PubMedGoogle Scholar
  56. Rozovskaia T, Tillib S, Smith S, Sedkov Y, Rozenblatt-Rosen O, Petruk S, Yano T, Nakamura T, Ben-Simchon L, Gildea J, Croce CM, Shearn A, Canaani E, Mazo A (1999) Trithorax and ASH1 interact directly and associate with the trithorax group-responsive bxd region of the Ultrabithorax promoter. Mol Cell Biol 19:6441–6447PubMedGoogle Scholar
  57. Shearn A (1989) The ash-1, ash-2 and trithorax genes of Drosophila melanogaster are functionally related. Genetics 121:517–525PubMedGoogle Scholar
  58. Simon JA, Tamkun JW (2002) Programming off and on states in chromatin: mechanisms of Polycomb and trithorax group complexes. Curr Opin Genet Dev 12:210–218PubMedCrossRefGoogle Scholar
  59. Sinclair DA, Clegg NJ, Antonchuk J, Milne TA, Stankunas K, Ruse C, Grigliatti TA, Kassis JA, Brock HW (1998) Enhancer of Polycomb is a suppressor of position-effect variegation in Drosophila melanogaster. Genetics 148:211–220PubMedGoogle Scholar
  60. Stassen MJ, Bailey D, Nelson S, Chinwalla V, Harte PJ (1995) The Drosophila trithorax proteins contain a novel variant of the nuclear receptor type DNA binding domain and an ancient conserved motif found in other chromosomal proteins. Mech Dev 52:209–223PubMedCrossRefGoogle Scholar
  61. Strutt H, Cavalli G, Paro R (1997) Co-localization of Polycomb protein and GAGA factor on regulatory elements responsible for the maintenance of homeotic gene expression. EMBO J 16:3621–3632PubMedCrossRefGoogle Scholar
  62. Torok T, Harvie PD, Buratovich M, Bryant PJ (1997) The product of proliferation disrupter is concentrated at centromeres and required for mitotic chromosome condensation and cell proliferation in Drosophila. Genes Dev 11:213–225PubMedCrossRefGoogle Scholar
  63. Tripoulas N, LaJeunesse D, Gildea J, Shearn A (1996) The Drosophila ash1 gene product, which is localized at specific sites on polytene chromosomes, contains a SET domain and a PHD finger. Genetics 143:913–928PubMedGoogle Scholar
  64. Tsukiyama T, Daniel C, Tamkun J, Wu C (1995) ISWI, a member of the SWI2/SNF2 ATPase family, encodes the 140 kDa subunit of the nucleosome remodeling factor. Cell 83:1021–1026PubMedCrossRefGoogle Scholar
  65. Weiler KS, Wakimoto BT (1995) Heterochromatin and gene expression in Drosophila. Annu Rev Genet 29:577–605PubMedCrossRefGoogle Scholar
  66. Zink D, Paro R (1989) In vivo binding pattern of trans-regulator of homeotic genes in Drosophila melanogaster. Nature 337:468–471PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Laura Fanti
    • 1
    Email author
  • Barbara Perrini
    • 1
  • Lucia Piacentini
    • 1
  • Maria Berloco
    • 2
  • Enzo Marchetti
    • 1
  • Gioacchino Palumbo
    • 2
  • Sergio Pimpinelli
    • 1
    Email author
  1. 1.Istituto Pasteur, Fondazione Cenci Bolognetti, Dipartimento di Genetica e Biologia molecolareUniversità “La Sapienza”RomeItaly
  2. 2.Dipartimento di Genetica e MicrobiologiaUniversità di BariBariItaly

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