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. 1995 Mar;15(3):1353–1363. doi: 10.1128/mcb.15.3.1353

Homotypic interactions of chicken GATA-1 can mediate transcriptional activation.

H Y Yang 1, T Evans 1
PMCID: PMC230359  PMID: 7862128

Abstract

We used a one-hybrid system to replace precisely the finger II chicken GATA-1 DNA-binding domain with the binding domain of bacterial repressor protein LexA. The LexA DNA-binding domain lacks amino acids that function for transcriptional activation, nuclear localization, or protein dimerization. This allowed us to analyze activities of GATA-1 sequences distinct from DNA binding. We found that strong transcriptional activating sequences that function independently of finger II are present in GATA-1. Sequences including finger I contain an independent nuclear localizing function. Our data are consistent with cooperative binding of two LexA-GATA-1 hybrid proteins on a palindromic operator. The sensitivity of our transcription assay provides the first evidence that GATA-1 can make homotypic interactions in vivo. The ability of a non-DNA-binding form of GATA-1 to activate gene expression by targeting to a bound GATA-1 derivative further supports the notion that GATA-1-GATA-1 interactions may have functional consequences. A coimmunoprecipitation assay was used to demonstrate that GATA-1 multimeric complexes form in solution by protein-protein interaction. The novel ability of GATA-1 to interact homotypically may be important for the formation of higher-order structures among distant regulatory elements that share binding sites for this transcription factor. We also used the system to test the ability of GATA-1 to interact heterotypically with other activators.

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Selected References

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  1. Abel T., Michelson A. M., Maniatis T. A Drosophila GATA family member that binds to Adh regulatory sequences is expressed in the developing fat body. Development. 1993 Nov;119(3):623–633. doi: 10.1242/dev.119.3.623. [DOI] [PubMed] [Google Scholar]
  2. Arceci R. J., King A. A., Simon M. C., Orkin S. H., Wilson D. B. Mouse GATA-4: a retinoic acid-inducible GATA-binding transcription factor expressed in endodermally derived tissues and heart. Mol Cell Biol. 1993 Apr;13(4):2235–2246. doi: 10.1128/mcb.13.4.2235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barton M. C., Madani N., Emerson B. M. The erythroid protein cGATA-1 functions with a stage-specific factor to activate transcription of chromatin-assembled beta-globin genes. Genes Dev. 1993 Sep;7(9):1796–1809. doi: 10.1101/gad.7.9.1796. [DOI] [PubMed] [Google Scholar]
  4. Brent R., Ptashne M. Mechanism of action of the lexA gene product. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4204–4208. doi: 10.1073/pnas.78.7.4204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang T. J., Scher B. M., Waxman S., Scher W. Inhibition of mouse GATA-1 function by the glucocorticoid receptor: possible mechanism of steroid inhibition of erythroleukemia cell differentiation. Mol Endocrinol. 1993 Apr;7(4):528–542. doi: 10.1210/mend.7.4.8502237. [DOI] [PubMed] [Google Scholar]
  6. Chiba T., Ikawa Y., Todokoro K. GATA-1 transactivates erythropoietin receptor gene, and erythropoietin receptor-mediated signals enhance GATA-1 gene expression. Nucleic Acids Res. 1991 Jul 25;19(14):3843–3848. doi: 10.1093/nar/19.14.3843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Choi O. R., Engel J. D. Developmental regulation of beta-globin gene switching. Cell. 1988 Oct 7;55(1):17–26. doi: 10.1016/0092-8674(88)90005-0. [DOI] [PubMed] [Google Scholar]
  8. Drevet J. R., Skeiky Y. A., Iatrou K. GATA-type zinc finger motif-containing sequences and chorion gene transcription factors of the silkworm Bombyx mori. J Biol Chem. 1994 Apr 8;269(14):10660–10667. [PubMed] [Google Scholar]
  9. Engel J. D. Developmental regulation of human beta-globin gene transcription: a switch of loyalties? Trends Genet. 1993 Sep;9(9):304–309. doi: 10.1016/0168-9525(93)90248-g. [DOI] [PubMed] [Google Scholar]
  10. Evans T., Felsenfeld G. The erythroid-specific transcription factor Eryf1: a new finger protein. Cell. 1989 Sep 8;58(5):877–885. doi: 10.1016/0092-8674(89)90940-9. [DOI] [PubMed] [Google Scholar]
  11. Evans T., Felsenfeld G. trans-Activation of a globin promoter in nonerythroid cells. Mol Cell Biol. 1991 Feb;11(2):843–853. doi: 10.1128/mcb.11.2.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Evans T., Reitman M., Felsenfeld G. An erythrocyte-specific DNA-binding factor recognizes a regulatory sequence common to all chicken globin genes. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5976–5980. doi: 10.1073/pnas.85.16.5976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Foley K. P., Engel J. D. Individual stage selector element mutations lead to reciprocal changes in beta- vs. epsilon-globin gene transcription: genetic confirmation of promoter competition during globin gene switching. Genes Dev. 1992 May;6(5):730–744. doi: 10.1101/gad.6.5.730. [DOI] [PubMed] [Google Scholar]
  14. Fong T. C., Emerson B. M. The erythroid-specific protein cGATA-1 mediates distal enhancer activity through a specialized beta-globin TATA box. Genes Dev. 1992 Apr;6(4):521–532. doi: 10.1101/gad.6.4.521. [DOI] [PubMed] [Google Scholar]
  15. Fu Y. H., Marzluf G. A. nit-2, the major positive-acting nitrogen regulatory gene of Neurospora crassa, encodes a sequence-specific DNA-binding protein. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5331–5335. doi: 10.1073/pnas.87.14.5331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Godowski P. J., Picard D., Yamamoto K. R. Signal transduction and transcriptional regulation by glucocorticoid receptor-LexA fusion proteins. Science. 1988 Aug 12;241(4867):812–816. doi: 10.1126/science.3043662. [DOI] [PubMed] [Google Scholar]
  17. Golemis E. A., Brent R. Fused protein domains inhibit DNA binding by LexA. Mol Cell Biol. 1992 Jul;12(7):3006–3014. doi: 10.1128/mcb.12.7.3006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gong Q., Dean A. Enhancer-dependent transcription of the epsilon-globin promoter requires promoter-bound GATA-1 and enhancer-bound AP-1/NF-E2. Mol Cell Biol. 1993 Feb;13(2):911–917. doi: 10.1128/mcb.13.2.911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ito E., Toki T., Ishihara H., Ohtani H., Gu L., Yokoyama M., Engel J. D., Yamamoto M. Erythroid transcription factor GATA-1 is abundantly transcribed in mouse testis. Nature. 1993 Apr 1;362(6419):466–468. doi: 10.1038/362466a0. [DOI] [PubMed] [Google Scholar]
  20. Jackson P. D., Evans T., Nickol J. M., Felsenfeld G. Developmental modulation of protein binding to beta-globin gene regulatory sites within chicken erythrocyte nuclei. Genes Dev. 1989 Dec;3(12A):1860–1873. doi: 10.1101/gad.3.12a.1860. [DOI] [PubMed] [Google Scholar]
  21. Kamens J., Brent R. A yeast transcription assay defines distinct rel and dorsal DNA recognition sequences. New Biol. 1991 Oct;3(10):1005–1013. [PubMed] [Google Scholar]
  22. Kelley C., Blumberg H., Zon L. I., Evans T. GATA-4 is a novel transcription factor expressed in endocardium of the developing heart. Development. 1993 Jul;118(3):817–827. doi: 10.1242/dev.118.3.817. [DOI] [PubMed] [Google Scholar]
  23. Kim B., Little J. W. Dimerization of a specific DNA-binding protein on the DNA. Science. 1992 Jan 10;255(5041):203–206. doi: 10.1126/science.1553548. [DOI] [PubMed] [Google Scholar]
  24. Knezetic J. A., Felsenfeld G. Identification and characterization of a chicken alpha-globin enhancer. Mol Cell Biol. 1989 Mar;9(3):893–901. doi: 10.1128/mcb.9.3.893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kudla B., Caddick M. X., Langdon T., Martinez-Rossi N. M., Bennett C. F., Sibley S., Davies R. W., Arst H. N., Jr The regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. Mutations affecting specificity of gene activation alter a loop residue of a putative zinc finger. EMBO J. 1990 May;9(5):1355–1364. doi: 10.1002/j.1460-2075.1990.tb08250.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Laverriere A. C., MacNeill C., Mueller C., Poelmann R. E., Burch J. B., Evans T. GATA-4/5/6, a subfamily of three transcription factors transcribed in developing heart and gut. J Biol Chem. 1994 Sep 16;269(37):23177–23184. [PubMed] [Google Scholar]
  27. Lech K., Anderson K., Brent R. DNA-bound Fos proteins activate transcription in yeast. Cell. 1988 Jan 29;52(2):179–184. doi: 10.1016/0092-8674(88)90506-5. [DOI] [PubMed] [Google Scholar]
  28. Lemarchandel V., Ghysdael J., Mignotte V., Rahuel C., Roméo P. H. GATA and Ets cis-acting sequences mediate megakaryocyte-specific expression. Mol Cell Biol. 1993 Jan;13(1):668–676. doi: 10.1128/mcb.13.1.668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Leonard M. W., Lim K. C., Engel J. D. Expression of the chicken GATA factor family during early erythroid development and differentiation. Development. 1993 Oct;119(2):519–531. doi: 10.1242/dev.119.2.519. [DOI] [PubMed] [Google Scholar]
  30. Leonard M., Brice M., Engel J. D., Papayannopoulou T. Dynamics of GATA transcription factor expression during erythroid differentiation. Blood. 1993 Aug 15;82(4):1071–1079. [PubMed] [Google Scholar]
  31. Liu F., Green M. R. A specific member of the ATF transcription factor family can mediate transcription activation by the adenovirus E1a protein. Cell. 1990 Jun 29;61(7):1217–1224. doi: 10.1016/0092-8674(90)90686-9. [DOI] [PubMed] [Google Scholar]
  32. Martin D. I., Orkin S. H. Transcriptional activation and DNA binding by the erythroid factor GF-1/NF-E1/Eryf 1. Genes Dev. 1990 Nov;4(11):1886–1898. doi: 10.1101/gad.4.11.1886. [DOI] [PubMed] [Google Scholar]
  33. Martin D. I., Tsai S. F., Orkin S. H. Increased gamma-globin expression in a nondeletion HPFH mediated by an erythroid-specific DNA-binding factor. Nature. 1989 Mar 30;338(6214):435–438. doi: 10.1038/338435a0. [DOI] [PubMed] [Google Scholar]
  34. Martin D. I., Zon L. I., Mutter G., Orkin S. H. Expression of an erythroid transcription factor in megakaryocytic and mast cell lineages. Nature. 1990 Mar 29;344(6265):444–447. doi: 10.1038/344444a0. [DOI] [PubMed] [Google Scholar]
  35. Martin K. J., Lillie J. W., Green M. R. Evidence for interaction of different eukaryotic transcriptional activators with distinct cellular targets. Nature. 1990 Jul 12;346(6280):147–152. doi: 10.1038/346147a0. [DOI] [PubMed] [Google Scholar]
  36. Matheny C., Day M. L., Milbrandt J. The nuclear localization signal of NGFI-A is located within the zinc finger DNA binding domain. J Biol Chem. 1994 Mar 18;269(11):8176–8181. [PubMed] [Google Scholar]
  37. Max-Audit I., Eleouet J. F., Roméo P. H. Transcriptional regulation of the pyruvate kinase erythroid-specific promoter. J Biol Chem. 1993 Mar 15;268(8):5431–5437. [PubMed] [Google Scholar]
  38. Oertel-Buchheit P., Schmidt-Dörr T., Granger-Schnarr M., Schnarr M. Spacing requirements between LexA operator half-sites can be relaxed by fusing the LexA DNA binding domain with some alternative dimerization domains. J Mol Biol. 1993 Jan 5;229(1):1–7. doi: 10.1006/jmbi.1993.1001. [DOI] [PubMed] [Google Scholar]
  39. Omichinski J. G., Clore G. M., Schaad O., Felsenfeld G., Trainor C., Appella E., Stahl S. J., Gronenborn A. M. NMR structure of a specific DNA complex of Zn-containing DNA binding domain of GATA-1. Science. 1993 Jul 23;261(5120):438–446. doi: 10.1126/science.8332909. [DOI] [PubMed] [Google Scholar]
  40. Omichinski J. G., Trainor C., Evans T., Gronenborn A. M., Clore G. M., Felsenfeld G. A small single-"finger" peptide from the erythroid transcription factor GATA-1 binds specifically to DNA as a zinc or iron complex. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1676–1680. doi: 10.1073/pnas.90.5.1676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Orkin S. H. GATA-binding transcription factors in hematopoietic cells. Blood. 1992 Aug 1;80(3):575–581. [PubMed] [Google Scholar]
  42. Orkin S. H. Globin gene regulation and switching: circa 1990. Cell. 1990 Nov 16;63(4):665–672. doi: 10.1016/0092-8674(90)90133-y. [DOI] [PubMed] [Google Scholar]
  43. Pascal E., Tjian R. Different activation domains of Sp1 govern formation of multimers and mediate transcriptional synergism. Genes Dev. 1991 Sep;5(9):1646–1656. doi: 10.1101/gad.5.9.1646. [DOI] [PubMed] [Google Scholar]
  44. Penner C. G., Davie J. R. Transcription factor GATA-1-multiprotein complexes and chicken erythroid development. FEBS Lett. 1994 Apr 11;342(3):273–277. doi: 10.1016/0014-5793(94)80515-6. [DOI] [PubMed] [Google Scholar]
  45. Pevny L., Simon M. C., Robertson E., Klein W. H., Tsai S. F., D'Agati V., Orkin S. H., Costantini F. Erythroid differentiation in chimaeric mice blocked by a targeted mutation in the gene for transcription factor GATA-1. Nature. 1991 Jan 17;349(6306):257–260. doi: 10.1038/349257a0. [DOI] [PubMed] [Google Scholar]
  46. Rahuel C., Vinit M. A., Lemarchandel V., Cartron J. P., Roméo P. H. Erythroid-specific activity of the glycophorin B promoter requires GATA-1 mediated displacement of a repressor. EMBO J. 1992 Nov;11(11):4095–4102. doi: 10.1002/j.1460-2075.1992.tb05502.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Ramain P., Heitzler P., Haenlin M., Simpson P. pannier, a negative regulator of achaete and scute in Drosophila, encodes a zinc finger protein with homology to the vertebrate transcription factor GATA-1. Development. 1993 Dec;119(4):1277–1291. doi: 10.1242/dev.119.4.1277. [DOI] [PubMed] [Google Scholar]
  48. Reitman M., Felsenfeld G. Mutational analysis of the chicken beta-globin enhancer reveals two positive-acting domains. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6267–6271. doi: 10.1073/pnas.85.17.6267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Schnarr M., Granger-Schnarr M., Hurstel S., Pouyet J. The carboxy-terminal domain of the LexA repressor oligomerises essentially as the entire protein. FEBS Lett. 1988 Jul 4;234(1):56–60. doi: 10.1016/0014-5793(88)81302-4. [DOI] [PubMed] [Google Scholar]
  50. Schnarr M., Oertel-Buchheit P., Kazmaier M., Granger-Schnarr M. DNA binding properties of the LexA repressor. Biochimie. 1991 Apr;73(4):423–431. doi: 10.1016/0300-9084(91)90109-e. [DOI] [PubMed] [Google Scholar]
  51. Schwartzbauer G., Schlesinger K., Evans T. Interaction of the erythroid transcription factor cGATA-1 with a critical auto-regulatory element. Nucleic Acids Res. 1992 Sep 11;20(17):4429–4436. doi: 10.1093/nar/20.17.4429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Simon M. C., Pevny L., Wiles M. V., Keller G., Costantini F., Orkin S. H. Rescue of erythroid development in gene targeted GATA-1- mouse embryonic stem cells. Nat Genet. 1992 May;1(2):92–98. doi: 10.1038/ng0592-92. [DOI] [PubMed] [Google Scholar]
  53. Spieth J., Shim Y. H., Lea K., Conrad R., Blumenthal T. elt-1, an embryonically expressed Caenorhabditis elegans gene homologous to the GATA transcription factor family. Mol Cell Biol. 1991 Sep;11(9):4651–4659. doi: 10.1128/mcb.11.9.4651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Sposi N. M., Zon L. I., Carè A., Valtieri M., Testa U., Gabbianelli M., Mariani G., Bottero L., Mather C., Orkin S. H. Cell cycle-dependent initiation and lineage-dependent abrogation of GATA-1 expression in pure differentiating hematopoietic progenitors. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6353–6357. doi: 10.1073/pnas.89.14.6353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Strauss E. C., Andrews N. C., Higgs D. R., Orkin S. H. In vivo footprinting of the human alpha-globin locus upstream regulatory element by guanine and adenine ligation-mediated polymerase chain reaction. Mol Cell Biol. 1992 May;12(5):2135–2142. doi: 10.1128/mcb.12.5.2135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Tamura S., Wang X. H., Maeda M., Futai M. Gastric DNA-binding proteins recognize upstream sequence motifs of parietal cell-specific genes. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10876–10880. doi: 10.1073/pnas.90.22.10876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Thliveris A. T., Little J. W., Mount D. W. Repression of the E coli recA gene requires at least two LexA protein monomers. Biochimie. 1991 Apr;73(4):449–456. doi: 10.1016/0300-9084(91)90112-e. [DOI] [PubMed] [Google Scholar]
  58. Tsai S. F., Martin D. I., Zon L. I., D'Andrea A. D., Wong G. G., Orkin S. H. Cloning of cDNA for the major DNA-binding protein of the erythroid lineage through expression in mammalian cells. Nature. 1989 Jun 8;339(6224):446–451. doi: 10.1038/339446a0. [DOI] [PubMed] [Google Scholar]
  59. Tsai S. F., Strauss E., Orkin S. H. Functional analysis and in vivo footprinting implicate the erythroid transcription factor GATA-1 as a positive regulator of its own promoter. Genes Dev. 1991 Jun;5(6):919–931. doi: 10.1101/gad.5.6.919. [DOI] [PubMed] [Google Scholar]
  60. Wall L., deBoer E., Grosveld F. The human beta-globin gene 3' enhancer contains multiple binding sites for an erythroid-specific protein. Genes Dev. 1988 Sep;2(9):1089–1100. doi: 10.1101/gad.2.9.1089. [DOI] [PubMed] [Google Scholar]
  61. Whyatt D. J., deBoer E., Grosveld F. The two zinc finger-like domains of GATA-1 have different DNA binding specificities. EMBO J. 1993 Dec 15;12(13):4993–5005. doi: 10.1002/j.1460-2075.1993.tb06193.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Winick J., Abel T., Leonard M. W., Michelson A. M., Chardon-Loriaux I., Holmgren R. A., Maniatis T., Engel J. D. A GATA family transcription factor is expressed along the embryonic dorsoventral axis in Drosophila melanogaster. Development. 1993 Dec;119(4):1055–1065. doi: 10.1242/dev.119.4.1055. [DOI] [PubMed] [Google Scholar]
  63. Yamamoto M., Ko L. J., Leonard M. W., Beug H., Orkin S. H., Engel J. D. Activity and tissue-specific expression of the transcription factor NF-E1 multigene family. Genes Dev. 1990 Oct;4(10):1650–1662. doi: 10.1101/gad.4.10.1650. [DOI] [PubMed] [Google Scholar]
  64. Yang H. Y., Evans T. Distinct roles for the two cGATA-1 finger domains. Mol Cell Biol. 1992 Oct;12(10):4562–4570. doi: 10.1128/mcb.12.10.4562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Yang Z., Gu L., Romeo P. H., Bories D., Motohashi H., Yamamoto M., Engel J. D. Human GATA-3 trans-activation, DNA-binding, and nuclear localization activities are organized into distinct structural domains. Mol Cell Biol. 1994 Mar;14(3):2201–2212. doi: 10.1128/mcb.14.3.2201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Ylikomi T., Bocquel M. T., Berry M., Gronemeyer H., Chambon P. Cooperation of proto-signals for nuclear accumulation of estrogen and progesterone receptors. EMBO J. 1992 Oct;11(10):3681–3694. doi: 10.1002/j.1460-2075.1992.tb05453.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Zhang C., Evans T. Differential regulation of the two xGATA-1 genes during Xenopus development. J Biol Chem. 1994 Jan 7;269(1):478–484. [PubMed] [Google Scholar]
  68. Zon L. I., Gurish M. F., Stevens R. L., Mather C., Reynolds D. S., Austen K. F., Orkin S. H. GATA-binding transcription factors in mast cells regulate the promoter of the mast cell carboxypeptidase A gene. J Biol Chem. 1991 Dec 5;266(34):22948–22953. [PubMed] [Google Scholar]
  69. Zon L. I., Mather C., Burgess S., Bolce M. E., Harland R. M., Orkin S. H. Expression of GATA-binding proteins during embryonic development in Xenopus laevis. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10642–10646. doi: 10.1073/pnas.88.23.10642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  70. Zon L. I., Yamaguchi Y., Yee K., Albee E. A., Kimura A., Bennett J. C., Orkin S. H., Ackerman S. J. Expression of mRNA for the GATA-binding proteins in human eosinophils and basophils: potential role in gene transcription. Blood. 1993 Jun 15;81(12):3234–3241. [PubMed] [Google Scholar]
  71. Zon L. I., Youssoufian H., Mather C., Lodish H. F., Orkin S. H. Activation of the erythropoietin receptor promoter by transcription factor GATA-1. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10638–10641. doi: 10.1073/pnas.88.23.10638. [DOI] [PMC free article] [PubMed] [Google Scholar]

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