Home LiteratureArticle Details
PMID: 2109322 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The second zinc-finger domain of poly(ADP-ribose) polymerase determines specificity for single-stranded breaks in DNA.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 8 ·1990-04-00 ·Pages 2990-4

Gradwohl G, Ménissier de Murcia JM, Molinete M, Simonin F, Koken M, Hoeijmakers JH, de Murcia G

Abstract

Poly(ADP-ribose) polymerase (EC 2.4.2.30) is a zinc-binding protein that specifically binds to a DNA strand break in a zinc-dependent manner. We describe here the cloning and expression in Escherichia coli of a cDNA fragment encoding the two putative zinc fingers (FI and FII) domain of the human poly(ADP-ribose) polymerase. Using site-directed mutagenesis, we identified the amino acids involved in metal coordination and analyzed the consequence of altering the proposed zinc-finger structures on DNA binding. Disruption of the metal binding ability of the second zinc finger, FII, dramatically reduced target DNA binding. In contrast, when the postulated Zn(II) ligands of FI were mutated, the DNA binding activity was only slightly affected. DNase I protection studies showed that the FII is involved in the specific recognition of a DNA strand break. These results demonstrate that poly(ADP-ribose) polymerase contains a type of zinc finger that differs from previously recognized classes in terms of both structure and function.

MeSH Terms
Amino Acid Sequence Base Sequence Cell Line Cloning, Molecular DNA, Single-Stranded/metabolism DNA-Binding Proteins/genetics,metabolism Escherichia coli/enzymology,genetics Humans Leukemia, Myelogenous, Chronic, BCR-ABL Positive Metalloproteins/genetics,metabolism Molecular Sequence Data Mutation Oligonucleotide Probes Poly(ADP-ribose) Polymerases/genetics,metabolism Protein Conformation Substrate Specificity Zinc/metabolism
Chemicals
DNA, Single-Stranded DNA-Binding Proteins Metalloproteins Oligonucleotide Probes Poly(ADP-ribose) Polymerases Zinc
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Gradwohl G
Institut de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Laboratoire de Biochimie II, Strasbourg, France.
Ménissier de Murcia J M
Molinete M
Simonin F
Koken M
Hoeijmakers J H
de Murcia G
References (28)
28 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Zinc-binding domain of poly(ADP-ribose)polymerase participates in the recognition of single strand breaks on DNA.
    J Mol Biol. 1989 Nov 5;210(1):229-33 PMID: 2511329
  3. A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase.
    J Mol Biol. 1975 May 25;94(3):441-8 PMID: 1100841
  4. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  5. Poly(ADP-ribose) synthesis in vitro programmed by damaged DNA. A comparison of DNA molecules containing different types of strand breaks.
    J Biol Chem. 1980 Nov 10;255(21):10502-8 PMID: 6253477
  6. High-level production of biologically active human alpha 1-antitrypsin in Escherichia coli.
    Proc Natl Acad Sci U S A. 1984 Feb;81(3):669-73 PMID: 6322161
  7. Poly(ADP-ribose) polymerase is a zinc metalloenzyme.
    Eur J Biochem. 1984 Aug 1;142(3):503-9 PMID: 6088227
  8. The primary structure of transcription factor TFIIIA has 12 consecutive repeats.
    FEBS Lett. 1985 Jul 8;186(2):271-4 PMID: 4007166
  9. Repetitive zinc-binding domains in the protein transcription factor IIIA from Xenopus oocytes.
    EMBO J. 1985 Jun;4(6):1609-14 PMID: 4040853
  10. Reconstitution and poly(ADP-ribosyl)ation of proteolytically fragmented poly(ADP-ribose) synthetase.
    J Biol Chem. 1986 Mar 15;261(8):3863-8 PMID: 3081511
  11. Modulation of chromatin superstructure induced by poly(ADP-ribose) synthesis and degradation.
    J Biol Chem. 1986 May 25;261(15):7011-7 PMID: 3084493
  12. TUF, the yeast DNA-binding factor specific for UASrpg upstream activating sequences: identification of the protein and its DNA-binding domain.
    Proc Natl Acad Sci U S A. 1987 Jun;84(11):3648-52 PMID: 3295867
  13. ADP-ribosylation of proteins. Enzymology and biological significance.
    Mol Biol Biochem Biophys. 1987;37:1-237 PMID: 3118181
  14. Primary structure of human poly(ADP-ribose) synthetase as deduced from cDNA sequence.
    J Biol Chem. 1987 Nov 25;262(33):15990-7 PMID: 2824474
  15. cDNA sequence, protein structure, and chromosomal location of the human gene for poly(ADP-ribose) polymerase.
    Proc Natl Acad Sci U S A. 1987 Dec;84(23):8370-4 PMID: 2891139
  16. Nucleotide sequence of a full-length cDNA for human fibroblast poly(ADP-ribose) polymerase.
    Biochem Biophys Res Commun. 1987 Oct 29;148(2):617-22 PMID: 3120710
  17. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.
    Science. 1988 Jan 29;239(4839):487-91 PMID: 2448875
  18. The steroid and thyroid hormone receptor superfamily.
    Science. 1988 May 13;240(4854):889-95 PMID: 3283939
  19. Structural and functional analysis of poly(ADP ribose) polymerase: an immunological study.
    Biochim Biophys Acta. 1988 Jul 13;950(2):147-60 PMID: 2454668
  20. The N-terminal DNA-binding 'zinc finger' of the oestrogen and glucocorticoid receptors determines target gene specificity.
    EMBO J. 1988 Oct;7(10):3037-44 PMID: 3141145
  21. Zinc-binding proteins detected by protein blotting.
    Anal Biochem. 1988 Jul;172(1):39-42 PMID: 3189772
  22. Metal binding 'finger' structures in the glucocorticoid receptor defined by site-directed mutagenesis.
    EMBO J. 1988 Aug;7(8):2503-8 PMID: 3191912
  23. Three amino acids of the oestrogen receptor are essential to its ability to distinguish an oestrogen from a glucocorticoid-responsive element.
    Nature. 1989 Mar 16;338(6212):271-4 PMID: 2922054
  24. Sequence and organization of the mouse poly (ADP-ribose) polymerase gene.
    Nucleic Acids Res. 1989 May 11;17(9):3387-401 PMID: 2498841
  25. Two amino acids within the knuckle of the first zinc finger specify DNA response element activation by the glucocorticoid receptor.
    Cell. 1989 Jun 30;57(7):1131-8 PMID: 2500250
  26. Determinants of target gene specificity for steroid/thyroid hormone receptors.
    Cell. 1989 Jun 30;57(7):1139-46 PMID: 2500251
  27. Poly(ADP-ribose)polymerase: a novel finger protein.
    Nucleic Acids Res. 1989 Jun 26;17(12):4689-98 PMID: 2501758
  28. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome.
    Blood. 1975 Mar;45(3):321-34 PMID: 163658
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.87.8.2990
Published
1990-04-00
Pages
2990-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC53819
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.87.8.2990 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