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

G. Gradwohl, J. Menissier De Murcia, M. Molinete, F. Simonin, M. Koken, J. H.J. Hoeijmakers, G. De Murcia

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250 Citaten (Scopus)

Samenvatting

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 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.

Originele taal-2Engels
Pagina's (van-tot)2990-2994
Aantal pagina's5
TijdschriftProceedings of the National Academy of Sciences of the United States of America
Volume87
Nummer van het tijdschrift8
DOI's
StatusGepubliceerd - 1990
Extern gepubliceerdJa

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