Expression in Escherichia coli of the 36 kDa domain of poly(ADP-ribose) polymerase and investigation of its DNA binding properties.
We have expressed in Escherichia coli the 36 kDa domain of the human poly(ADP-ribose) polymerase. This polypeptide comprises the C-terminal part of the DNA binding domain, as well as the automodification region of the enzyme, but lacks the zinc-finger motifs of the N-terminal region and the C-terminal catalytic domain. By probing the crude E. coli protein extracts with radioactive DNA probes (South-Western blots), we have shown that the 36 kDa domain binds a DNA probe of 222 bp but does not bind a shorter probe of 66 bp. This interaction is stronger when the polypeptide is fused to the 55 kDa catalytic domain of the enzyme.
Thibodeau
J
J
Laboratorie du métabolisme du poly(ADP-ribose), Endocrinologie moléculaire, Centre Hospitalier de l'Université Laval, Québec, Canada.
Potvin
F
F
Kirkland
J B
JB
Poirier
G
G
D000595:Amino Acid Sequence; D004247:DNA / Q000378:metabolism; D015342:DNA Probes; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics* / Q000378:metabolism
D015342:DNA Probes; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases