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PMID: 2159531 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

In vitro transcriptional activation, dimerization, and DNA-binding specificity of the Epstein-Barr virus Zta protein.

Journal of virology ·Vol. 64 ·No. 6 ·1990-06-00 ·Pages 2560-8

Lieberman PM, Berk AJ

Abstract

The Epstein-Barr virus BZLF1 immediate-early gene encodes a transcriptional activator protein, Zta, which acts as a key regulatory switch in the transition between the latent and lytic viral life cycle. In this work, full-length Zta was expressed at high levels in Escherichia coli and purified to homogeneity by DNA affinity chromatography. The bacterial protein bound to specific target sequences (Zta response elements) and activated transcription in vitro from an Epstein-Barr virus early target promoter (BHLF1). Zta bound to DNA as a dimer. The formation of a heterodimer with a Zta deletion mutant was detected by gel electrophoresis mobility shift assays. Footprinting analysis on the BHLF1, BZLF1, and simian virus 40 control regions revealed multiple binding sites with no simple consensus sequence. Zta bound upstream from its own promoter at low concentrations, while at high concentrations it bound at the transcription start site, suggesting that it may activate and then autoregulate its own expression. These results demonstrate that Zta is a sequence-specific DNA-binding transcription factor.

MeSH Terms
Amino Acid Sequence Base Sequence Chromatography, Affinity Cloning, Molecular DNA-Binding Proteins/genetics Deoxyribonuclease I Electrophoresis, Polyacrylamide Gel Escherichia coli/genetics Herpesvirus 4, Human/genetics,metabolism Macromolecular Substances Molecular Sequence Data Molecular Weight Oligonucleotide Probes Recombinant Proteins/isolation & purification,metabolism Trans-Activators/genetics,isolation & purification,metabolism Transcription, Genetic Transcriptional Activation Viral Proteins
Chemicals
BZLF1 protein, Herpesvirus 4, Human DNA-Binding Proteins Macromolecular Substances Oligonucleotide Probes Recombinant Proteins Trans-Activators Viral Proteins Deoxyribonuclease I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Lieberman P M
Molecular Biology Institute, University of California, Los Angeles 90024.
Berk A J
References (63)
63 references, click to expand
  1. trans activation of the latent Epstein-Barr virus (EBV) genome after transfection of the EBV DNA fragment.
    J Virol. 1986 Mar;57(3):1016-22 PMID: 3005608
  2. The spliced BZLF1 gene of Epstein-Barr virus (EBV) transactivates an early EBV promoter and induces the virus productive cycle.
    J Virol. 1989 Jul;63(7):3109-16 PMID: 2542618
  3. Calcium modulation activates Epstein-Barr virus genome in latently infected cells.
    Science. 1986 Jun 20;232(4757):1554-6 PMID: 3012779
  4. Definitive identification of a member of the Epstein-Barr virus nuclear protein 3 family.
    Proc Natl Acad Sci U S A. 1986 Aug;83(15):5693-7 PMID: 3016714
  5. Affinity purification of sequence-specific DNA binding proteins.
    Proc Natl Acad Sci U S A. 1986 Aug;83(16):5889-93 PMID: 3461465
  6. An Epstein-Barr virus (EBV)-determined nuclear antigen (EBNA5) partly encoded by the transformation-associated Bam WYH region of EBV DNA: preferential expression in lymphoblastoid cell lines.
    Proc Natl Acad Sci U S A. 1986 Sep;83(17):6641-5 PMID: 3018741
  7. Persisting oncogenic herpesvirus induced by the tumour promotor TPA.
    Nature. 1978 Mar 23;272(5651):373-5 PMID: 204874
  8. Induction of the Epstein-Barr virus (EBV) cycle in latently infected cells by n-butyrate.
    Virology. 1979 Apr 15;94(1):228-31 PMID: 220786
  9. Two small RNAs encoded by Epstein-Barr virus and complexed with protein are precipitated by antibodies from patients with systemic lupus erythematosus.
    Proc Natl Acad Sci U S A. 1981 Feb;78(2):805-9 PMID: 6262773
  10. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
    Anal Biochem. 1981 Apr;112(2):195-203 PMID: 6266278
  11. A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system.
    Nucleic Acids Res. 1981 Jul 10;9(13):3047-60 PMID: 6269071
  12. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis.
    Nucleic Acids Res. 1981 Dec 11;9(23):6505-25 PMID: 6275366
  13. Silver staining of proteins in polyacrylamide gels.
    Anal Biochem. 1981 Nov 15;118(1):197-203 PMID: 6175245
  14. Characterization of the major Epstein-Barr virus-specific RNA in Burkitt lymphoma-derived cells.
    J Virol. 1982 Feb;41(2):376-89 PMID: 6281455
  15. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  16. SV40 T antigen binding site mutations that affect autoregulation.
    Cell. 1983 Apr;32(4):1227-40 PMID: 6301686
  17. Periodic features in the amino acid sequence of nematode myosin rod.
    J Mol Biol. 1983 Mar 15;164(4):605-26 PMID: 6341606
  18. Organization of the Epstein-Barr virus DNA molecule. III. Location of the P3HR-1 deletion junction and characterization of the NotI repeat units that form part of the template for an abundant 12-O-tetradecanoylphorbol-13-acetate-induced mRNA transcript.
    J Virol. 1983 Oct;48(1):135-48 PMID: 6310141
  19. The use of pKc30 and its derivatives for controlled expression of genes.
    Methods Enzymol. 1983;101:123-38 PMID: 6310317
  20. One of two Epstein-Barr virus nuclear antigens contains a glycine-alanine copolymer domain.
    Proc Natl Acad Sci U S A. 1983 Sep;80(18):5665-9 PMID: 6310587
  21. Epstein-Barr virus with heterogeneous DNA disrupts latency.
    J Virol. 1984 Apr;50(1):174-82 PMID: 6321789
  22. Structure of the 5' ends of immunoglobulin genes: a novel conserved sequence.
    Proc Natl Acad Sci U S A. 1984 May;81(9):2650-4 PMID: 6425835
  23. Correct transcription of an immunoglobulin kappa gene requires an upstream fragment containing conserved sequence elements.
    Nature. 1984 Jul 5-11;310(5972):71-4 PMID: 6330567
  24. Nucleotide sequence of an mRNA transcribed in latent growth-transforming virus infection indicates that it may encode a membrane protein.
    J Virol. 1984 Aug;51(2):411-9 PMID: 6086953
  25. A membrane protein encoded by Epstein-Barr virus in latent growth-transforming infection.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):7207-11 PMID: 6095274
  26. Purification and characterization of transcription factor IIIC2.
    J Biol Chem. 1989 Jun 25;264(18):10726-31 PMID: 2732244
  27. The Epstein-Barr virus early protein EB1 activates transcription from different responsive elements including AP-1 binding sites.
    EMBO J. 1989 May;8(5):1447-53 PMID: 2548844
  28. Epstein-Barr virus bicistronic mRNAs generated by facultative splicing code for two transcriptional trans-activators.
    EMBO J. 1989 Jun;8(6):1819-26 PMID: 2548857
  29. Sp1 activates transcription without enhancing DNA-binding activity of the TATA box factor.
    Mol Cell Biol. 1989 Aug;9(8):3299-307 PMID: 2677669
  30. Scissors-grip model for DNA recognition by a family of leucine zipper proteins.
    Science. 1989 Nov 17;246(4932):911-6 PMID: 2683088
  31. The zta transactivator involved in induction of lytic cycle gene expression in Epstein-Barr virus-infected lymphocytes binds to both AP-1 and ZRE sites in target promoter and enhancer regions.
    J Virol. 1990 Mar;64(3):1143-55 PMID: 2154599
  32. Two related but differentially expressed potential membrane proteins encoded by the EcoRI Dhet region of Epstein-Barr virus B95-8.
    J Virol. 1985 Feb;53(2):528-35 PMID: 2982035
  33. A second nuclear protein is encoded by Epstein-Barr virus in latent infection.
    Science. 1985 Mar 8;227(4691):1238-40 PMID: 2983420
  34. Activation of expression of latent Epstein-Barr herpesvirus after gene transfer with a small cloned subfragment of heterogeneous viral DNA.
    Proc Natl Acad Sci U S A. 1985 Jun;82(12):4085-9 PMID: 2987963
  35. Two distinct transcription factors bind to the HSV thymidine kinase promoter in vitro.
    Cell. 1985 Sep;42(2):559-72 PMID: 2992804
  36. A third viral nuclear protein in lymphoblasts immortalized by Epstein-Barr virus.
    Proc Natl Acad Sci U S A. 1985 Sep;82(17):5944-8 PMID: 2994055
  37. Sequence-specific DNA binding of the Epstein-Barr virus nuclear antigen (EBNA-1) to clustered sites in the plasmid maintenance region.
    Cell. 1985 Oct;42(3):859-68 PMID: 2996781
  38. Structure and evolution of two related transcription units of Epstein-Barr virus carrying small tandem repeats.
    J Virol. 1985 Dec;56(3):987-95 PMID: 2999451
  39. Homologous recognition of a promoter domain common to the MSV LTR and the HSV tk gene.
    Cell. 1986 Feb 28;44(4):565-76 PMID: 3004739
  40. Both Epstein-Barr virus (EBV)-encoded trans-acting factors, EB1 and EB2, are required to activate transcription from an EBV early promoter.
    EMBO J. 1986 Dec 1;5(12):3243-9 PMID: 3028777
  41. A TATA box implicated in E1A transcriptional activation of a simple adenovirus 2 promoter.
    Nature. 1987 Apr 2-8;326(6112):512-5 PMID: 2951598
  42. Epstein-Barr virus gene expression in P3HR1-superinfected Raji cells.
    J Virol. 1987 Oct;61(10):3120-32 PMID: 3041034
  43. Mapping of the gene coding for Epstein-Barr virus-determined nuclear antigen EBNA3 and its transient overexpression in a human cell line by using an adenovirus expression vector.
    J Virol. 1987 Oct;61(10):3340-4 PMID: 3041055
  44. GCN4, a eukaryotic transcriptional activator protein, binds as a dimer to target DNA.
    EMBO J. 1987 Sep;6(9):2781-4 PMID: 3678204
  45. Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain.
    Cell. 1987 Dec 24;51(6):1079-90 PMID: 3319186
  46. A fifth Epstein-Barr virus nuclear protein (EBNA3C) is expressed in latently infected growth-transformed lymphocytes.
    J Virol. 1988 Apr;62(4):1330-8 PMID: 2831394
  47. Latency comes of age for herpesviruses.
    Cell. 1988 Mar 25;52(6):787-9 PMID: 2832064
  48. A sixth Epstein-Barr virus nuclear protein (EBNA3B) is expressed in latently infected growth-transformed lymphocytes.
    J Virol. 1988 Jun;62(6):2173-8 PMID: 2835517
  49. A new Epstein-Barr virus transactivator, R, induces expression of a cytoplasmic early antigen.
    J Virol. 1988 Jul;62(7):2274-84 PMID: 2836611
  50. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins.
    Science. 1988 Jun 24;240(4860):1759-64 PMID: 3289117
  51. A spliced Epstein-Barr virus gene expressed in immortalized lymphocytes is created by circularization of the linear viral genome.
    EMBO J. 1988 Mar;7(3):769-74 PMID: 2840285
  52. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity.
    Nucleic Acids Res. 1978 Sep;5(9):3157-70 PMID: 212715
  53. Direct correlation between a negative autoregulatory response element at the cap site of the herpes simplex virus type 1 IE175 (alpha 4) promoter and a specific binding site for the IE175 (ICP4) protein.
    J Virol. 1988 Nov;62(11):4307-20 PMID: 2845144
  54. The role of the leucine zipper in the fos-jun interaction.
    Nature. 1988 Dec 15;336(6200):646-51 PMID: 2974122
  55. The Epstein-Barr virus (EBV) early promoter DR contains a cis-acting element responsive to the EBV transactivator EB1 and an enhancer with constitutive and inducible activities.
    J Virol. 1989 Feb;63(2):607-14 PMID: 2536096
  56. Evidence that the leucine zipper is a coiled coil.
    Science. 1989 Jan 27;243(4890):538-42 PMID: 2911757
  57. The Epstein-Barr virus (EBV) BZLF1 immediate-early gene product differentially affects latent versus productive EBV promoters.
    J Virol. 1989 Apr;63(4):1729-36 PMID: 2538653
  58. The DNA binding domain of the rat liver nuclear protein C/EBP is bipartite.
    Science. 1989 Mar 31;243(4899):1681-8 PMID: 2494700
  59. Leucine repeats and an adjacent DNA binding domain mediate the formation of functional cFos-cJun heterodimers.
    Science. 1989 Mar 31;243(4899):1689-94 PMID: 2494701
  60. Parallel association of Fos and Jun leucine zippers juxtaposes DNA binding domains.
    Science. 1989 Mar 31;243(4899):1695-9 PMID: 2494702
  61. Epstein-Barr virus BZLF1 trans-activator specifically binds to a consensus AP-1 site and is related to c-fos.
    EMBO J. 1989 Jan;8(1):127-32 PMID: 2540954
  62. Responsiveness of the Epstein-Barr virus NotI repeat promoter to the Z transactivator is mediated in a cell-type-specific manner by two independent signal regions.
    J Virol. 1989 Jul;63(7):3040-50 PMID: 2542612
  63. Four virally determined nuclear antigens are expressed in Epstein-Barr virus-transformed cells.
    Proc Natl Acad Sci U S A. 1986 Mar;83(5):1499-503 PMID: 3485288
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.64.6.2560-2568.1990
Published
1990-06-00
Pages
2560-8
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC249432
Subset
IM
Grants
NCI NIH HHS · CA25237 · United States
External Links
PubMed source DOI: 10.1128/JVI.64.6.2560-2568.1990 PMC Full Text View journal details
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