Home LiteratureArticle Details
PMID: 1835093 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Conversion of the E1A Cys4 zinc finger to a nonfunctional His2,Cys2 zinc finger by a single point mutation.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 22 ·1991-11-15 ·Pages 9989-93

Webster LC, Zhang K, Chance B, Ayene I, Culp JS, Huang WJ, Wu FY, Ricciardi RP

Abstract

Trans-activation by the adenovirus E1A 289R protein requires a zinc finger defined by Cys-154, Cys-157, Cys-171, and Cys-174. Whereas individually replacing the four cysteine residues with serines resulted in a loss of transactivation, only three of the Cys----Ser mutants (C157S, C171S, and C174S) lost the ability to bind Zn(II). X-ray absorption fine structure analysis revealed that, in the wild-type protein, Zn(II) is coordinated by four cysteine residues whereas in the C154S mutant, Zn(II) is coordinated by two histidines and two cysteines. The mutant protein probably retains, as ligands, two cysteines on the right side of the zinc finger (Cys-171 and Cys-174) and recruits two of the four histidines on the left side (His-149, His-152, His-158, and His-160), despite the presence of Cys-157. This finding may shed light on the general structural requirements of zinc fingers.

MeSH Terms
Adenoviridae/genetics Adenovirus Early Proteins Amino Acid Sequence Base Sequence Binding Sites Cloning, Molecular Codon/genetics Cysteine DNA-Binding Proteins/genetics Escherichia coli/genetics HeLa Cells Humans Molecular Sequence Data Mutagenesis, Site-Directed Oncogene Proteins, Viral/genetics Plasmids Protein Conformation Transcriptional Activation Transfection Zinc/metabolism Zinc Fingers/genetics
Chemicals
Adenovirus Early Proteins Codon DNA-Binding Proteins Oncogene Proteins, Viral Zinc Cysteine
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Webster L C
Institute for Structural Functional Studies, Philadelphia, PA 19104.
Zhang K
Chance B
Ayene I
Culp J S
Huang W J
Wu F Y
Ricciardi R P
References (43)
43 references, click to expand
  1. DNA binding specificity of steroid receptors.
    Cell. 1989 Jun 30;57(7):1065-8 PMID: 2661016
  2. Multiple transcription factor binding sites mediate adenovirus E1A transactivation.
    J Virol. 1989 Aug;63(8):3499-506 PMID: 2545919
  3. 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
  4. Genetic evidence that zinc is an essential co-factor in the DNA binding domain of GAL4 protein.
    Nature. 1987 Jul 23-29;328(6128):353-5 PMID: 3299106
  5. HeLa cell beta-tubulin gene transcription is stimulated by adenovirus 5 in parallel with viral early genes by an E1a-dependent mechanism.
    Mol Cell Biol. 1984 Dec;4(12):2792-801 PMID: 6527689
  6. Affinity purification of sequence-specific DNA binding proteins.
    Proc Natl Acad Sci U S A. 1986 Aug;83(16):5889-93 PMID: 3461465
  7. EXAFS study of the zinc-binding sites in the protein transcription factor IIIA.
    Nature. 1986 Dec 18-31;324(6098):698-9 PMID: 3796733
  8. Repetitive zinc-binding domains in the protein transcription factor IIIA from Xenopus oocytes.
    EMBO J. 1985 Jun;4(6):1609-14 PMID: 4040853
  9. Solution structure of the glucocorticoid receptor DNA-binding domain.
    Science. 1990 Jul 13;249(4965):157-60 PMID: 2115209
  10. Solution structure of a zinc finger domain of yeast ADR1.
    Proteins. 1990;7(3):215-26 PMID: 2114025
  11. A specific member of the ATF transcription factor family can mediate transcription activation by the adenovirus E1a protein.
    Cell. 1990 Jun 29;61(7):1217-24 PMID: 2142019
  12. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.
    Mol Cell Biol. 1982 Sep;2(9):1044-51 PMID: 6960240
  13. Regulation of adenovirus transcription by an E1a gene in microinjected Xenopus laevis oocytes.
    Mol Cell Biol. 1983 Dec;3(12):2131-42 PMID: 6318084
  14. Xenopus transcription factor A requires zinc for binding to the 5 S RNA gene.
    J Biol Chem. 1983 Dec 10;258(23):14120-5 PMID: 6196359
  15. Specific interaction of a purified transcription factor with an internal control region of 5S RNA genes.
    Cell. 1980 Mar;19(3):717-28 PMID: 6153931
  16. DNA sequences bound specifically by glucocorticoid receptor in vitro render a heterologous promoter hormone responsive in vivo.
    Cell. 1983 Jun;33(2):489-99 PMID: 6190571
  17. Adenovirus E1a gene product expressed at high levels in Escherichia coli is functional.
    Science. 1984 Jun 22;224(4655):1343-6 PMID: 6374895
  18. Transcriptional activation and subsequent control of the human heat shock gene during adenovirus infection.
    Mol Cell Biol. 1983 Nov;3(11):2058-65 PMID: 6689197
  19. E1A control of gene expression is mediated by sequences 5' to the transcriptional starts of the early viral genes.
    Mol Cell Biol. 1983 Jul;3(7):1222-34 PMID: 6888379
  20. An x-ray absorption study of the iron site in bacterial photosynthetic reaction centers.
    Biophys J. 1982 Feb;37(2):539-51 PMID: 6977382
  21. Expression of early adenovirus genes requires a viral encoded acidic polypeptide.
    Proc Natl Acad Sci U S A. 1981 Oct;78(10):6121-5 PMID: 6947217
  22. Resolving the functions of overlapping viral genes by site-specific mutagenesis at a mRNA splice site.
    Nature. 1982 Feb 4;295(5848):380-4 PMID: 7057903
  23. An adenovirus type 5 early gene function regulates expression of other early viral genes.
    Proc Natl Acad Sci U S A. 1979 Aug;76(8):3665-9 PMID: 291030
  24. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  25. Pre-early adenovirus 5 gene product regulates synthesis of early viral messenger RNAs.
    Cell. 1979 Aug;17(4):935-44 PMID: 487437
  26. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  27. Three-dimensional solution structure of a single zinc finger DNA-binding domain.
    Science. 1989 Aug 11;245(4918):635-7 PMID: 2503871
  28. Zinc-dependent structure of a single-finger domain of yeast ADR1.
    Science. 1988 Sep 16;241(4872):1489-92 PMID: 3047872
  29. Cysteine residues in the zinc finger and amino acids adjacent to the finger are necessary for DNA binding by the LAC9 regulatory protein of Kluyveromyces lactis.
    Mol Cell Biol. 1988 Sep;8(9):3726-33 PMID: 3146691
  30. Disruption of a putative Cys-zinc interaction eliminates the biological activity of the Krüppel finger protein.
    Nature. 1988 Mar 3;332(6159):90-2 PMID: 3126398
  31. Adenovirus 5 early region 1A host range mutants hr3, hr4, and hr5 contain point mutations which generate single amino acid substitutions.
    J Virol. 1985 Oct;56(1):66-74 PMID: 3861875
  32. Potential metal-binding domains in nucleic acid binding proteins.
    Science. 1986 Apr 25;232(4749):485-7 PMID: 2421409
  33. Zinc-finger motifs expressed in E. coli and folded in vitro direct specific binding to DNA.
    Nature. 1988 Mar 17;332(6161):284-6 PMID: 2831463
  34. Adenovirus promoters and E1A transactivation.
    Annu Rev Genet. 1986;20:45-79 PMID: 3028247
  35. The 289-amino acid E1A protein of adenovirus binds zinc in a region that is important for trans-activation.
    Proc Natl Acad Sci U S A. 1988 Sep;85(17):6450-4 PMID: 2970640
  36. DNA-binding properties of an adenovirus 289R E1A protein.
    EMBO J. 1988 Mar;7(3):835-41 PMID: 2969333
  37. Identification of a cellular transcription factor involved in E1A trans-activation.
    Cell. 1986 Apr 25;45(2):219-28 PMID: 2938741
  38. 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
  39. An adenovirus type 5 E1A protein with a single amino acid substitution blocks wild-type E1A transactivation.
    Mol Cell Biol. 1987 Mar;7(3):1004-11 PMID: 2951587
  40. Zinc fingers: gilt by association.
    Cell. 1988 Jan 15;52(1):1-3 PMID: 3125980
  41. The yeast regulatory protein ADR1 binds in a zinc-dependent manner to the upstream activating sequence of ADH2.
    Mol Cell Biol. 1988 Oct;8(10):4552-6 PMID: 3141794
  42. The function and structure of the metal coordination sites within the glucocorticoid receptor DNA binding domain.
    Nature. 1988 Aug 11;334(6182):543-6 PMID: 3043231
  43. The active site of hemerythrin as determined by X-ray absorption fine structure.
    Biochemistry. 1988 Sep 20;27(19):7470-9 PMID: 3207685
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.88.22.9989
Published
1991-11-15
Pages
9989-93
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC52852
Subset
IM
Grants
NCI NIH HHS · CA-09171 · United States
NCI NIH HHS · CA-29797 · United States
NIGMS NIH HHS · GM-28057 · United States
External Links
PubMed source DOI: 10.1073/pnas.88.22.9989 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2