Skip to main content
PMC home page
Genetics logoLink to Genetics
. 2004 Jul;167(3):1213–1223. doi: 10.1534/genetics.103.018721

The direct interaction between ASH2, a Drosophila trithorax group protein, and SKTL, a nuclear phosphatidylinositol 4-phosphate 5-kinase, implies a role for phosphatidylinositol 4,5-bisphosphate in maintaining transcriptionally active chromatin.

Mimi K Cheng 1, Allen Shearn 1
PMCID: PMC1470965  PMID: 15280236

Abstract

The products of trithorax group (trxG) genes maintain active transcription of many important developmental regulatory genes, including homeotic genes. Several trxG proteins have been shown to act in multimeric protein complexes that modify chromatin structure. ASH2, the product of the Drosophila trxG gene absent, small, or homeotic discs 2 (ash2) is a component of a 500-kD complex. In this article, we provide biochemical evidence that ASH2 binds directly to Skittles (SKTL), a predicted phosphatidylinositol 4-phosphate 5-kinase, and genetic evidence that the association of these proteins is functionally significant. We also show that histone H1 hyperphosphorylation is dramatically increased in both ash2 and sktl mutant polytene chromosomes. These results suggest that ASH2 maintains active transcription by binding a producer of nuclear phosphoinositides and downregulating histone H1 hyperphosphorylation.

Full Text

The Full Text of this article is available as a PDF (563.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aasland R., Gibson T. J., Stewart A. F. The PHD finger: implications for chromatin-mediated transcriptional regulation. Trends Biochem Sci. 1995 Feb;20(2):56–59. doi: 10.1016/s0968-0004(00)88957-4. [DOI] [PubMed] [Google Scholar]
  2. Adams M. D., Celniker S. E., Holt R. A., Evans C. A., Gocayne J. D., Amanatides P. G., Scherer S. E., Li P. W., Hoskins R. A., Galle R. F. The genome sequence of Drosophila melanogaster. Science. 2000 Mar 24;287(5461):2185–2195. doi: 10.1126/science.287.5461.2185. [DOI] [PubMed] [Google Scholar]
  3. Adamson A. L., Shearn A. Molecular genetic analysis of Drosophila ash2, a member of the trithorax group required for imaginal disc pattern formation. Genetics. 1996 Oct;144(2):621–633. doi: 10.1093/genetics/144.2.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Akam M. The molecular basis for metameric pattern in the Drosophila embryo. Development. 1987 Sep;101(1):1–22. [PubMed] [Google Scholar]
  5. Anderson R. A., Boronenkov I. V., Doughman S. D., Kunz J., Loijens J. C. Phosphatidylinositol phosphate kinases, a multifaceted family of signaling enzymes. J Biol Chem. 1999 Apr 9;274(15):9907–9910. doi: 10.1074/jbc.274.15.9907. [DOI] [PubMed] [Google Scholar]
  6. Banks G. C., Deterding L. J., Tomer K. B., Archer T. K. Hormone-mediated dephosphorylation of specific histone H1 isoforms. J Biol Chem. 2001 Jul 30;276(39):36467–36473. doi: 10.1074/jbc.M104641200. [DOI] [PubMed] [Google Scholar]
  7. Beisel Christian, Imhof Axel, Greene Jaime, Kremmer Elisabeth, Sauer Frank. Histone methylation by the Drosophila epigenetic transcriptional regulator Ash1. Nature. 2002 Oct 9;419(6909):857–862. doi: 10.1038/nature01126. [DOI] [PubMed] [Google Scholar]
  8. Beltran Sergi, Blanco Enrique, Serras Florenci, Pérez-Villamil Beatriz, Guigó Roderic, Artavanis-Tsakonas Spyros, Corominas Montserrat. Transcriptional network controlled by the trithorax-group gene ash2 in Drosophila melanogaster. Proc Natl Acad Sci U S A. 2003 Mar 7;100(6):3293–3298. doi: 10.1073/pnas.0538075100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Boronenkov I. V., Loijens J. C., Umeda M., Anderson R. A. Phosphoinositide signaling pathways in nuclei are associated with nuclear speckles containing pre-mRNA processing factors. Mol Biol Cell. 1998 Dec;9(12):3547–3560. doi: 10.1091/mbc.9.12.3547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Bresnick E. H., Bustin M., Marsaud V., Richard-Foy H., Hager G. L. The transcriptionally-active MMTV promoter is depleted of histone H1. Nucleic Acids Res. 1992 Jan 25;20(2):273–278. doi: 10.1093/nar/20.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chong L. D., Traynor-Kaplan A., Bokoch G. M., Schwartz M. A. The small GTP-binding protein Rho regulates a phosphatidylinositol 4-phosphate 5-kinase in mammalian cells. Cell. 1994 Nov 4;79(3):507–513. doi: 10.1016/0092-8674(94)90259-3. [DOI] [PubMed] [Google Scholar]
  12. Cochet C., Filhol O., Payrastre B., Hunter T., Gill G. N. Interaction between the epidermal growth factor receptor and phosphoinositide kinases. J Biol Chem. 1991 Jan 5;266(1):637–644. [PubMed] [Google Scholar]
  13. Croston G. E., Kerrigan L. A., Lira L. M., Marshak D. R., Kadonaga J. T. Sequence-specific antirepression of histone H1-mediated inhibition of basal RNA polymerase II transcription. Science. 1991 Feb 8;251(4994):643–649. doi: 10.1126/science.1899487. [DOI] [PubMed] [Google Scholar]
  14. Dedon P. C., Soults J. A., Allis C. D., Gorovsky M. A. Formaldehyde cross-linking and immunoprecipitation demonstrate developmental changes in H1 association with transcriptionally active genes. Mol Cell Biol. 1991 Mar;11(3):1729–1733. doi: 10.1128/mcb.11.3.1729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Divecha N., Banfić H., Irvine R. F. Inositides and the nucleus and inositides in the nucleus. Cell. 1993 Aug 13;74(3):405–407. doi: 10.1016/0092-8674(93)80041-c. [DOI] [PubMed] [Google Scholar]
  16. Dou Y., Gorovsky M. A. Phosphorylation of linker histone H1 regulates gene expression in vivo by creating a charge patch. Mol Cell. 2000 Aug;6(2):225–231. doi: 10.1016/s1097-2765(00)00024-1. [DOI] [PubMed] [Google Scholar]
  17. Dou Y., Mizzen C. A., Abrams M., Allis C. D., Gorovsky M. A. Phosphorylation of linker histone H1 regulates gene expression in vivo by mimicking H1 removal. Mol Cell. 1999 Oct;4(4):641–647. doi: 10.1016/s1097-2765(00)80215-4. [DOI] [PubMed] [Google Scholar]
  18. Duncan I. The bithorax complex. Annu Rev Genet. 1987;21:285–319. doi: 10.1146/annurev.ge.21.120187.001441. [DOI] [PubMed] [Google Scholar]
  19. Francis N. J., Kingston R. E. Mechanisms of transcriptional memory. Nat Rev Mol Cell Biol. 2001 Jun;2(6):409–421. doi: 10.1038/35073039. [DOI] [PubMed] [Google Scholar]
  20. Gildea J. J., Lopez R., Shearn A. A screen for new trithorax group genes identified little imaginal discs, the Drosophila melanogaster homologue of human retinoblastoma binding protein 2. Genetics. 2000 Oct;156(2):645–663. doi: 10.1093/genetics/156.2.645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gozani Or, Karuman Philip, Jones David R., Ivanov Dmitri, Cha James, Lugovskoy Alexey A., Baird Cheryl L., Zhu Hong, Field Seth J., Lessnick Stephen L. The PHD finger of the chromatin-associated protein ING2 functions as a nuclear phosphoinositide receptor. Cell. 2003 Jul 11;114(1):99–111. doi: 10.1016/s0092-8674(03)00480-x. [DOI] [PubMed] [Google Scholar]
  22. Guo K., Nichol R., Skehel P., Dormann D., Weijer C. J., Williams J. G., Pears C. A Dictyostelium nuclear phosphatidylinositol phosphate kinase required for developmental gene expression. EMBO J. 2001 Nov 1;20(21):6017–6027. doi: 10.1093/emboj/20.21.6017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hay J. C., Fisette P. L., Jenkins G. H., Fukami K., Takenawa T., Anderson R. A., Martin T. F. ATP-dependent inositide phosphorylation required for Ca(2+)-activated secretion. Nature. 1995 Mar 9;374(6518):173–177. doi: 10.1038/374173a0. [DOI] [PubMed] [Google Scholar]
  24. Hohmann P. Phosphorylation of H1 histones. Mol Cell Biochem. 1983;57(1):81–92. doi: 10.1007/BF00223526. [DOI] [PubMed] [Google Scholar]
  25. Horn Peter J., Carruthers Lenny M., Logie Colin, Hill David A., Solomon Mark J., Wade Paul A., Imbalzano Anthony N., Hansen Jeffrey C., Peterson Craig L. Phosphorylation of linker histones regulates ATP-dependent chromatin remodeling enzymes. Nat Struct Biol. 2002 Apr;9(4):263–267. doi: 10.1038/nsb776. [DOI] [PubMed] [Google Scholar]
  26. Ikegawa S., Isomura M., Koshizuka Y., Nakamura Y. Cloning and characterization of ASH2L and Ash2l, human and mouse homologs of the Drosophila ash2 gene. Cytogenet Cell Genet. 1999;84(3-4):167–172. doi: 10.1159/000015248. [DOI] [PubMed] [Google Scholar]
  27. Ishihara H., Shibasaki Y., Kizuki N., Katagiri H., Yazaki Y., Asano T., Oka Y. Cloning of cDNAs encoding two isoforms of 68-kDa type I phosphatidylinositol-4-phosphate 5-kinase. J Biol Chem. 1996 Sep 27;271(39):23611–23614. doi: 10.1074/jbc.271.39.23611. [DOI] [PubMed] [Google Scholar]
  28. Janmey P. A. Phosphoinositides and calcium as regulators of cellular actin assembly and disassembly. Annu Rev Physiol. 1994;56:169–191. doi: 10.1146/annurev.ph.56.030194.001125. [DOI] [PubMed] [Google Scholar]
  29. LaJeunesse D., Shearn A. Trans-regulation of thoracic homeotic selector genes of the Antennapedia and bithorax complexes by the trithorax group genes: absent, small, and homeotic discs 1 and 2. Mech Dev. 1995 Sep;53(1):123–139. doi: 10.1016/0925-4773(95)00430-0. [DOI] [PubMed] [Google Scholar]
  30. Laport Ginna G., Levine Bruce L., Stadtmauer Edward A., Schuster Stephen J., Luger Selina M., Grupp Stephan, Bunin Nancy, Strobl Frank J., Cotte Julio, Zheng Zhaohui. Adoptive transfer of costimulated T cells induces lymphocytosis in patients with relapsed/refractory non-Hodgkin lymphoma following CD34+-selected hematopoietic cell transplantation. Blood. 2003 May 22;102(6):2004–2013. doi: 10.1182/blood-2003-01-0095. [DOI] [PubMed] [Google Scholar]
  31. Laybourn P. J., Kadonaga J. T. Role of nucleosomal cores and histone H1 in regulation of transcription by RNA polymerase II. Science. 1991 Oct 11;254(5029):238–245. doi: 10.1126/science.254.5029.238. [DOI] [PubMed] [Google Scholar]
  32. Manzoli F. A., Maraldi N. M., Cocco L., Capitani S., Facchini A. Chromatin phospholipids in normal and chronic lymphocytic leukemia lymphocytes. Cancer Res. 1977 Mar;37(3):843–849. [PubMed] [Google Scholar]
  33. Miller T., Krogan N. J., Dover J., Erdjument-Bromage H., Tempst P., Johnston M., Greenblatt J. F., Shilatifard A. COMPASS: a complex of proteins associated with a trithorax-related SET domain protein. Proc Natl Acad Sci U S A. 2001 Oct 30;98(23):12902–12907. doi: 10.1073/pnas.231473398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nagy Peter L., Griesenbeck Joachim, Kornberg Roger D., Cleary Michael L. A trithorax-group complex purified from Saccharomyces cerevisiae is required for methylation of histone H3. Proc Natl Acad Sci U S A. 2001 Dec 18;99(1):90–94. doi: 10.1073/pnas.221596698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Odom A. R., Stahlberg A., Wente S. R., York J. D. A role for nuclear inositol 1,4,5-trisphosphate kinase in transcriptional control. Science. 2000 Mar 17;287(5460):2026–2029. doi: 10.1126/science.287.5460.2026. [DOI] [PubMed] [Google Scholar]
  36. Osborne S. L., Thomas C. L., Gschmeissner S., Schiavo G. Nuclear PtdIns(4,5)P2 assembles in a mitotically regulated particle involved in pre-mRNA splicing. J Cell Sci. 2001 Jul;114(Pt 13):2501–2511. doi: 10.1242/jcs.114.13.2501. [DOI] [PubMed] [Google Scholar]
  37. Papoulas O., Beek S. J., Moseley S. L., McCallum C. M., Sarte M., Shearn A., Tamkun J. W. The Drosophila trithorax group proteins BRM, ASH1 and ASH2 are subunits of distinct protein complexes. Development. 1998 Oct;125(20):3955–3966. doi: 10.1242/dev.125.20.3955. [DOI] [PubMed] [Google Scholar]
  38. Payrastre B., Nievers M., Boonstra J., Breton M., Verkleij A. J., Van Bergen en Henegouwen P. M. A differential location of phosphoinositide kinases, diacylglycerol kinase, and phospholipase C in the nuclear matrix. J Biol Chem. 1992 Mar 15;267(8):5078–5084. [PubMed] [Google Scholar]
  39. Perry C. A., Annunziato A. T. Influence of histone acetylation on the solubility, H1 content and DNase I sensitivity of newly assembled chromatin. Nucleic Acids Res. 1989 Jun 12;17(11):4275–4291. doi: 10.1093/nar/17.11.4275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ponting C., Schultz J., Bork P. SPRY domains in ryanodine receptors (Ca(2+)-release channels). Trends Biochem Sci. 1997 Jun;22(6):193–194. doi: 10.1016/s0968-0004(97)01049-9. [DOI] [PubMed] [Google Scholar]
  41. Rana R. S., Hokin L. E. Role of phosphoinositides in transmembrane signaling. Physiol Rev. 1990 Jan;70(1):115–164. doi: 10.1152/physrev.1990.70.1.115. [DOI] [PubMed] [Google Scholar]
  42. Ren X. D., Bokoch G. M., Traynor-Kaplan A., Jenkins G. H., Anderson R. A., Schwartz M. A. Physical association of the small GTPase Rho with a 68-kDa phosphatidylinositol 4-phosphate 5-kinase in Swiss 3T3 cells. Mol Biol Cell. 1996 Mar;7(3):435–442. doi: 10.1091/mbc.7.3.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ridsdale J. A., Hendzel M. J., Delcuve G. P., Davie J. R. Histone acetylation alters the capacity of the H1 histones to condense transcriptionally active/competent chromatin. J Biol Chem. 1990 Mar 25;265(9):5150–5156. [PubMed] [Google Scholar]
  44. Robbins J., Dilworth S. M., Laskey R. A., Dingwall C. Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell. 1991 Feb 8;64(3):615–623. doi: 10.1016/0092-8674(91)90245-t. [DOI] [PubMed] [Google Scholar]
  45. Rogers S., Wells R., Rechsteiner M. Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science. 1986 Oct 17;234(4774):364–368. doi: 10.1126/science.2876518. [DOI] [PubMed] [Google Scholar]
  46. Roguev A., Schaft D., Shevchenko A., Pijnappel W. W., Wilm M., Aasland R., Stewart A. F. The Saccharomyces cerevisiae Set1 complex includes an Ash2 homologue and methylates histone 3 lysine 4. EMBO J. 2001 Dec 17;20(24):7137–7148. doi: 10.1093/emboj/20.24.7137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Roguev Assen, Schaft Daniel, Shevchenko Anna, Aasland Rein, Shevchenko Andrej, Stewart A. Francis. High conservation of the Set1/Rad6 axis of histone 3 lysine 4 methylation in budding and fission yeasts. J Biol Chem. 2002 Dec 17;278(10):8487–8493. doi: 10.1074/jbc.M209562200. [DOI] [PubMed] [Google Scholar]
  48. Roth S. Y., Allis C. D. Chromatin condensation: does histone H1 dephosphorylation play a role? Trends Biochem Sci. 1992 Mar;17(3):93–98. doi: 10.1016/0968-0004(92)90243-3. [DOI] [PubMed] [Google Scholar]
  49. Shearn A. The ash-1, ash-2 and trithorax genes of Drosophila melanogaster are functionally related. Genetics. 1989 Mar;121(3):517–525. doi: 10.1093/genetics/121.3.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Shen Xuetong, Xiao Hua, Ranallo Ryan, Wu Wei-Hua, Wu Carl. Modulation of ATP-dependent chromatin-remodeling complexes by inositol polyphosphates. Science. 2002 Nov 14;299(5603):112–114. doi: 10.1126/science.1078068. [DOI] [PubMed] [Google Scholar]
  51. Simon J. Locking in stable states of gene expression: transcriptional control during Drosophila development. Curr Opin Cell Biol. 1995 Jun;7(3):376–385. doi: 10.1016/0955-0674(95)80093-x. [DOI] [PubMed] [Google Scholar]
  52. Steger David J., Haswell Elizabeth S., Miller Aimee L., Wente Susan R., O'Shea Erin K. Regulation of chromatin remodeling by inositol polyphosphates. Science. 2002 Nov 14;299(5603):114–116. doi: 10.1126/science.1078062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Toker A., Meyer M., Reddy K. K., Falck J. R., Aneja R., Aneja S., Parra A., Burns D. J., Ballas L. M., Cantley L. C. Activation of protein kinase C family members by the novel polyphosphoinositides PtdIns-3,4-P2 and PtdIns-3,4,5-P3. J Biol Chem. 1994 Dec 23;269(51):32358–32367. [PubMed] [Google Scholar]
  54. Toker A. The synthesis and cellular roles of phosphatidylinositol 4,5-bisphosphate. Curr Opin Cell Biol. 1998 Apr;10(2):254–261. doi: 10.1016/s0955-0674(98)80148-8. [DOI] [PubMed] [Google Scholar]
  55. Tripoulas N., LaJeunesse D., Gildea J., Shearn A. The Drosophila ash1 gene product, which is localized at specific sites on polytene chromosomes, contains a SET domain and a PHD finger. Genetics. 1996 Jun;143(2):913–928. doi: 10.1093/genetics/143.2.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. White R. A., Wilcox M. Protein products of the bithorax complex in Drosophila. Cell. 1984 Nov;39(1):163–171. doi: 10.1016/0092-8674(84)90202-2. [DOI] [PubMed] [Google Scholar]
  57. Wysocka Joanna, Myers Michael P., Laherty Carol D., Eisenman Robert N., Herr Winship. Human Sin3 deacetylase and trithorax-related Set1/Ash2 histone H3-K4 methyltransferase are tethered together selectively by the cell-proliferation factor HCF-1. Genes Dev. 2003 Apr 1;17(7):896–911. doi: 10.1101/gad.252103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Yamamoto A., DeWald D. B., Boronenkov I. V., Anderson R. A., Emr S. D., Koshland D. Novel PI(4)P 5-kinase homologue, Fab1p, essential for normal vacuole function and morphology in yeast. Mol Biol Cell. 1995 May;6(5):525–539. doi: 10.1091/mbc.6.5.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. York J. D., Odom A. R., Murphy R., Ives E. B., Wente S. R. A phospholipase C-dependent inositol polyphosphate kinase pathway required for efficient messenger RNA export. Science. 1999 Jul 2;285(5424):96–100. doi: 10.1126/science.285.5424.96. [DOI] [PubMed] [Google Scholar]
  60. Yu H., Fukami K., Watanabe Y., Ozaki C., Takenawa T. Phosphatidylinositol 4,5-bisphosphate reverses the inhibition of RNA transcription caused by histone H1. Eur J Biochem. 1998 Jan 15;251(1-2):281–287. doi: 10.1046/j.1432-1327.1998.2510281.x. [DOI] [PubMed] [Google Scholar]
  61. Zhao K., Wang W., Rando O. J., Xue Y., Swiderek K., Kuo A., Crabtree G. R. Rapid and phosphoinositol-dependent binding of the SWI/SNF-like BAF complex to chromatin after T lymphocyte receptor signaling. Cell. 1998 Nov 25;95(5):625–636. doi: 10.1016/s0092-8674(00)81633-5. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES