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. 1995 Aug 1;14(15):3777–3787. doi: 10.1002/j.1460-2075.1995.tb00047.x

Staf, a novel zinc finger protein that activates the RNA polymerase III promoter of the selenocysteine tRNA gene.

C Schuster 1, E Myslinski 1, A Krol 1, P Carbon 1
PMCID: PMC394452  PMID: 7641696

Abstract

The selenocysteine tRNA gene (tRNA(Sec)) is atypical. Though transcribed by RNA polymerase III like all other tRNA genes, its basal promoter elements are distinct and reside essentially upstream of the coding region. In addition, transcription from the basal promoter is activated by a 15 bp activator element. In this report we describe the cloning and functional characterization of Staf (selenocysteine tRNA gene transcription activating factor), a novel Xenopus laevis transcription factor which binds to the tRNA(Sec) activator element and mediates its activation properties. The 600 amino acid Staf protein contains seven zinc fingers and a separate acidic activation domain. Seven highly conserved regions were detected between Staf and human ZNF76, a protein of unknown function, thereby aiding in predicting the locations of the functional domains of Staf. With the use of a novel expression assay in X.laevis oocytes we succeeded in demonstrating that Staf can activate the RNA polymerase III promoter of the tRNA(Sec) gene. This constitutes the first demonstration of the capacity of a cloned factor to activate RNA polymerase III transcription in vivo.

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Selected References

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

  1. Behne D., Hilmert H., Scheid S., Gessner H., Elger W. Evidence for specific selenium target tissues and new biologically important selenoproteins. Biochim Biophys Acta. 1988 Jul 14;966(1):12–21. doi: 10.1016/0304-4165(88)90123-7. [DOI] [PubMed] [Google Scholar]
  2. Bellefroid E. J., Poncelet D. A., Lecocq P. J., Revelant O., Martial J. A. The evolutionarily conserved Krüppel-associated box domain defines a subfamily of eukaryotic multifingered proteins. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3608–3612. doi: 10.1073/pnas.88.9.3608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradley L. C., Snape A., Bhatt S., Wilkinson D. G. The structure and expression of the Xenopus Krox-20 gene: conserved and divergent patterns of expression in rhombomeres and neural crest. Mech Dev. 1993 Jan;40(1-2):73–84. doi: 10.1016/0925-4773(93)90089-g. [DOI] [PubMed] [Google Scholar]
  4. Brown R. S., Sander C., Argos P. The primary structure of transcription factor TFIIIA has 12 consecutive repeats. FEBS Lett. 1985 Jul 8;186(2):271–274. doi: 10.1016/0014-5793(85)80723-7. [DOI] [PubMed] [Google Scholar]
  5. Caput D., Beutler B., Hartog K., Thayer R., Brown-Shimer S., Cerami A. Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1670–1674. doi: 10.1073/pnas.83.6.1670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carbon P., Krol A. Transcription of the Xenopus laevis selenocysteine tRNA(Ser)Sec gene: a system that combines an internal B box and upstream elements also found in U6 snRNA genes. EMBO J. 1991 Mar;10(3):599–606. doi: 10.1002/j.1460-2075.1991.tb07987.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carbon P., Murgo S., Ebel J. P., Krol A., Tebb G., Mattaj L. W. A common octamer motif binding protein is involved in the transcription of U6 snRNA by RNA polymerase III and U2 snRNA by RNA polymerase II. Cell. 1987 Oct 9;51(1):71–79. doi: 10.1016/0092-8674(87)90011-0. [DOI] [PubMed] [Google Scholar]
  8. Carey M. Mechanistic advances in eukaryotic gene activation. Curr Opin Cell Biol. 1991 Jun;3(3):452–460. doi: 10.1016/0955-0674(91)90073-8. [DOI] [PubMed] [Google Scholar]
  9. Chavrier P., Janssen-Timmen U., Mattéi M. G., Zerial M., Bravo R., Charnay P. Structure, chromosome location, and expression of the mouse zinc finger gene Krox-20: multiple gene products and coregulation with the proto-oncogene c-fos. Mol Cell Biol. 1989 Feb;9(2):787–797. doi: 10.1128/mcb.9.2.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chavrier P., Vesque C., Galliot B., Vigneron M., Dollé P., Duboule D., Charnay P. The segment-specific gene Krox-20 encodes a transcription factor with binding sites in the promoter region of the Hox-1.4 gene. EMBO J. 1990 Apr;9(4):1209–1218. doi: 10.1002/j.1460-2075.1990.tb08228.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Courey A. J., Tjian R. Analysis of Sp1 in vivo reveals multiple transcriptional domains, including a novel glutamine-rich activation motif. Cell. 1988 Dec 2;55(5):887–898. doi: 10.1016/0092-8674(88)90144-4. [DOI] [PubMed] [Google Scholar]
  12. Danzeiser D. A., Urso O., Kunkel G. R. Functional characterization of elements in a human U6 small nuclear RNA gene distal control region. Mol Cell Biol. 1993 Aug;13(8):4670–4678. doi: 10.1128/mcb.13.8.4670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Delwel R., Funabiki T., Kreider B. L., Morishita K., Ihle J. N. Four of the seven zinc fingers of the Evi-1 myeloid-transforming gene are required for sequence-specific binding to GA(C/T)AAGA(T/C)AAGATAA. Mol Cell Biol. 1993 Jul;13(7):4291–4300. doi: 10.1128/mcb.13.7.4291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gabrielsen O. S., Sentenac A. RNA polymerase III (C) and its transcription factors. Trends Biochem Sci. 1991 Nov;16(11):412–416. doi: 10.1016/0968-0004(91)90166-s. [DOI] [PubMed] [Google Scholar]
  15. Green S., Issemann I., Sheer E. A versatile in vivo and in vitro eukaryotic expression vector for protein engineering. Nucleic Acids Res. 1988 Jan 11;16(1):369–369. doi: 10.1093/nar/16.1.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Herbomel P., Bourachot B., Yaniv M. Two distinct enhancers with different cell specificities coexist in the regulatory region of polyoma. Cell. 1984 Dec;39(3 Pt 2):653–662. doi: 10.1016/0092-8674(84)90472-0. [DOI] [PubMed] [Google Scholar]
  17. Keller A. D., Maniatis T. Only two of the five zinc fingers of the eukaryotic transcriptional repressor PRDI-BF1 are required for sequence-specific DNA binding. Mol Cell Biol. 1992 May;12(5):1940–1949. doi: 10.1128/mcb.12.5.1940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kleinert H., Bredow S., Benecke B. J. Expression of a human 7S K RNA gene in vivo requires a novel pol III upstream element. EMBO J. 1990 Mar;9(3):711–718. doi: 10.1002/j.1460-2075.1990.tb08164.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Klocke B., Köster M., Hille S., Bouwmeester T., Böhm S., Pieler T., Knöchel W. The FAR domain defines a new Xenopus laevis zinc finger protein subfamily with specific RNA homopolymer binding activity. Biochim Biophys Acta. 1994 Jan 18;1217(1):81–89. [PubMed] [Google Scholar]
  20. Knöchel W., Pöting A., Köster M., el Baradi T., Nietfeld W., Bouwmeester T., Pieler T. Evolutionary conserved modules associated with zinc fingers in Xenopus laevis. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6097–6100. doi: 10.1073/pnas.86.16.6097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Krasnow M. A., Saffman E. E., Kornfeld K., Hogness D. S. Transcriptional activation and repression by Ultrabithorax proteins in cultured Drosophila cells. Cell. 1989 Jun 16;57(6):1031–1043. doi: 10.1016/0092-8674(89)90341-3. [DOI] [PubMed] [Google Scholar]
  22. Krieg P. A., Melton D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 1984 Sep 25;12(18):7057–7070. doi: 10.1093/nar/12.18.7057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Krol A., Carbon P., Ebel J. P., Appel B. Xenopus tropicalis U6 snRNA genes transcribed by Pol III contain the upstream promoter elements used by Pol II dependent U snRNA genes. Nucleic Acids Res. 1987 Mar 25;15(6):2463–2478. doi: 10.1093/nar/15.6.2463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kunkel G. R., Pederson T. Upstream elements required for efficient transcription of a human U6 RNA gene resemble those of U1 and U2 genes even though a different polymerase is used. Genes Dev. 1988 Feb;2(2):196–204. doi: 10.1101/gad.2.2.196. [DOI] [PubMed] [Google Scholar]
  25. Kunkel G. R. RNA polymerase III transcription of genes that lack internal control regions. Biochim Biophys Acta. 1991 Jan 17;1088(1):1–9. doi: 10.1016/0167-4781(91)90146-d. [DOI] [PubMed] [Google Scholar]
  26. Lemaire P., Garrett N., Gurdon J. B. Expression cloning of Siamois, a Xenopus homeobox gene expressed in dorsal-vegetal cells of blastulae and able to induce a complete secondary axis. Cell. 1995 Apr 7;81(1):85–94. doi: 10.1016/0092-8674(95)90373-9. [DOI] [PubMed] [Google Scholar]
  27. Lescure A., Murgo S., Carbon P., Krol A. The proximal promoter and the start site cooperate to specify correct U1 snRNA transcription initiation by RNA polymerase II. Nucleic Acids Res. 1992 Apr 11;20(7):1573–1578. doi: 10.1093/nar/20.7.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lescure A., Tebb G., Mattaj I. W., Krol A., Carbon P. A factor with Sp1 DNA-binding specificity stimulates Xenopus U6 snRNA in vivo transcription by RNA polymerase III. J Mol Biol. 1992 Nov 20;228(2):387–394. doi: 10.1016/0022-2836(92)90828-8. [DOI] [PubMed] [Google Scholar]
  29. Lobo S. M., Hernandez N. A 7 bp mutation converts a human RNA polymerase II snRNA promoter into an RNA polymerase III promoter. Cell. 1989 Jul 14;58(1):55–67. doi: 10.1016/0092-8674(89)90402-9. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Mattaj I. W., Dathan N. A., Parry H. D., Carbon P., Krol A. Changing the RNA polymerase specificity of U snRNA gene promoters. Cell. 1988 Nov 4;55(3):435–442. doi: 10.1016/0092-8674(88)90029-3. [DOI] [PubMed] [Google Scholar]
  32. Meissner W., Wanandi I., Carbon P., Krol A., Seifart K. H. Transcription factors required for the expression of Xenopus laevis selenocysteine tRNA in vitro. Nucleic Acids Res. 1994 Feb 25;22(4):553–559. doi: 10.1093/nar/22.4.553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Miller J., McLachlan A. D., Klug A. Repetitive zinc-binding domains in the protein transcription factor IIIA from Xenopus oocytes. EMBO J. 1985 Jun;4(6):1609–1614. doi: 10.1002/j.1460-2075.1985.tb03825.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mitchell P. J., Tjian R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989 Jul 28;245(4916):371–378. doi: 10.1126/science.2667136. [DOI] [PubMed] [Google Scholar]
  35. Murphy S., Pierani A., Scheidereit C., Melli M., Roeder R. G. Purified octamer binding transcription factors stimulate RNA polymerase III--mediated transcription of the 7SK RNA gene. Cell. 1989 Dec 22;59(6):1071–1080. doi: 10.1016/0092-8674(89)90763-0. [DOI] [PubMed] [Google Scholar]
  36. Murphy S., Yoon J. B., Gerster T., Roeder R. G. Oct-1 and Oct-2 potentiate functional interactions of a transcription factor with the proximal sequence element of small nuclear RNA genes. Mol Cell Biol. 1992 Jul;12(7):3247–3261. doi: 10.1128/mcb.12.7.3247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Myslinski E., Krol A., Carbon P. Optimal tRNA((Ser)Sec) gene activity requires an upstream SPH motif. Nucleic Acids Res. 1992 Jan 25;20(2):203–209. doi: 10.1093/nar/20.2.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Myslinski E., Schuster C., Huet J., Sentenac A., Krol A., Carbon P. Point mutations 5' to the tRNA selenocysteine TATA box alter RNA polymerase III transcription by affecting the binding of TBP. Nucleic Acids Res. 1993 Dec 25;21(25):5852–5858. doi: 10.1093/nar/21.25.5852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Myslinski E., Schuster C., Krol A., Carbon P. Promoter strength and structure dictate module composition in RNA polymerase III transcriptional activator elements. J Mol Biol. 1993 Nov 20;234(2):311–318. doi: 10.1006/jmbi.1993.1588. [DOI] [PubMed] [Google Scholar]
  40. Numoto M., Niwa O., Kaplan J., Wong K. K., Merrell K., Kamiya K., Yanagihara K., Calame K. Transcriptional repressor ZF5 identifies a new conserved domain in zinc finger proteins. Nucleic Acids Res. 1993 Aug 11;21(16):3767–3775. doi: 10.1093/nar/21.16.3767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Pavletich N. P., Pabo C. O. Crystal structure of a five-finger GLI-DNA complex: new perspectives on zinc fingers. Science. 1993 Sep 24;261(5129):1701–1707. doi: 10.1126/science.8378770. [DOI] [PubMed] [Google Scholar]
  42. Pazin M. J., Kamakaka R. T., Kadonaga J. T. ATP-dependent nucleosome reconfiguration and transcriptional activation from preassembled chromatin templates. Science. 1994 Dec 23;266(5193):2007–2011. doi: 10.1126/science.7801129. [DOI] [PubMed] [Google Scholar]
  43. Perkins N. D., Agranoff A. B., Pascal E., Nabel G. J. An interaction between the DNA-binding domains of RelA(p65) and Sp1 mediates human immunodeficiency virus gene activation. Mol Cell Biol. 1994 Oct;14(10):6570–6583. doi: 10.1128/mcb.14.10.6570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Ragoussis J., Senger G., Mockridge I., Sanseau P., Ruddy S., Dudley K., Sheer D., Trowsdale J. A testis-expressed Zn finger gene (ZNF76) in human 6p21.3 centromeric to the MHC is closely linked to the human homolog of the t-complex gene tcp-11. Genomics. 1992 Nov;14(3):673–679. doi: 10.1016/s0888-7543(05)80167-3. [DOI] [PubMed] [Google Scholar]
  45. Simmen K. A., Bernués J., Parry H. D., Stunnenberg H. G., Berkenstam A., Cavallini B., Egly J. M., Mattaj I. W. TFIID is required for in vitro transcription of the human U6 gene by RNA polymerase III. EMBO J. 1991 Jul;10(7):1853–1862. doi: 10.1002/j.1460-2075.1991.tb07711.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Singh H., LeBowitz J. H., Baldwin A. S., Jr, Sharp P. A. Molecular cloning of an enhancer binding protein: isolation by screening of an expression library with a recognition site DNA. Cell. 1988 Feb 12;52(3):415–423. doi: 10.1016/s0092-8674(88)80034-5. [DOI] [PubMed] [Google Scholar]
  47. Stadtman T. C. Biosynthesis and function of selenocysteine-containing enzymes. J Biol Chem. 1991 Sep 5;266(25):16257–16260. [PubMed] [Google Scholar]
  48. Staudt L. M., Clerc R. G., Singh H., LeBowitz J. H., Sharp P. A., Baltimore D. Cloning of a lymphoid-specific cDNA encoding a protein binding the regulatory octamer DNA motif. Science. 1988 Jul 29;241(4865):577–580. doi: 10.1126/science.3399892. [DOI] [PubMed] [Google Scholar]
  49. Sturm R. A., Das G., Herr W. The ubiquitous octamer-binding protein Oct-1 contains a POU domain with a homeo box subdomain. Genes Dev. 1988 Dec;2(12A):1582–1599. doi: 10.1101/gad.2.12a.1582. [DOI] [PubMed] [Google Scholar]
  50. Tanaka M., Lai J. S., Herr W. Promoter-selective activation domains in Oct-1 and Oct-2 direct differential activation of an snRNA and mRNA promoter. Cell. 1992 Feb 21;68(4):755–767. doi: 10.1016/0092-8674(92)90150-b. [DOI] [PubMed] [Google Scholar]
  51. Thisse C., Perrin-Schmitt F., Stoetzel C., Thisse B. Sequence-specific transactivation of the Drosophila twist gene by the dorsal gene product. Cell. 1991 Jun 28;65(7):1191–1201. doi: 10.1016/0092-8674(91)90014-p. [DOI] [PubMed] [Google Scholar]
  52. Tjian R., Maniatis T. Transcriptional activation: a complex puzzle with few easy pieces. Cell. 1994 Apr 8;77(1):5–8. doi: 10.1016/0092-8674(94)90227-5. [DOI] [PubMed] [Google Scholar]
  53. Vesque C., Charnay P. Mapping functional regions of the segment-specific transcription factor Krox-20. Nucleic Acids Res. 1992 May 25;20(10):2485–2492. doi: 10.1093/nar/20.10.2485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Vinson C. R., LaMarco K. L., Johnson P. F., Landschulz W. H., McKnight S. L. In situ detection of sequence-specific DNA binding activity specified by a recombinant bacteriophage. Genes Dev. 1988 Jul;2(7):801–806. doi: 10.1101/gad.2.7.801. [DOI] [PubMed] [Google Scholar]
  55. Waldschmidt R., Wanandi I., Seifart K. H. Identification of transcription factors required for the expression of mammalian U6 genes in vitro. EMBO J. 1991 Sep;10(9):2595–2603. doi: 10.1002/j.1460-2075.1991.tb07801.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Webster N., Jin J. R., Green S., Hollis M., Chambon P. The yeast UASG is a transcriptional enhancer in human HeLa cells in the presence of the GAL4 trans-activator. Cell. 1988 Jan 29;52(2):169–178. doi: 10.1016/0092-8674(88)90505-3. [DOI] [PubMed] [Google Scholar]
  57. Willis I. M. RNA polymerase III. Genes, factors and transcriptional specificity. Eur J Biochem. 1993 Feb 15;212(1):1–11. doi: 10.1111/j.1432-1033.1993.tb17626.x. [DOI] [PubMed] [Google Scholar]

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