Poly(ADP-ribose) polymerase-1: association with nuclear lamins in rodent liver cells.
The distribution of poly(ADP-ribose) polymerase-1 (PARP-1) over different nuclear compartments was studied by nuclear fractionation procedures and Western analysis revealing a prominent role of the nuclear matrix. This structure is operationally defined by the solubility properties of the A- and B-type lamins under defined experimental conditions. We consistently observed that most of the nuclear matrix-associated PARP-1 partitioned, in an active form, with the insoluble, lamin-enriched protein fractions that were prepared by a variety of established biochemical procedures. These PARP-1-protein interactions resisted salt extraction, disulfide reduction, RNase and DNase digestion. An inherent ability of PARP-1 to reassemble with the lamins became evident after a cycle of solubilization/dialysis using either urea or Triton X-100 and disulfide reduction, indicating that these interactions were dominated by hydrophobic forces. Together with in vivo crosslinking and co-immunoprecipitation experiments our results show that the lamins are prominent PARP-1-binding partners which could contribute to the functional sequestration of the enzyme on the nuclear matrix.
Vidaković
Melita
M
Molecular Biology Laboratory, Institute for Biological Research, 29th November 142, 11060 Belgrade, Serbia and Montenegro, Yugoslavia.
Grdović
Nevena
N
Quesada
Piera
P
Bode
Jürgen
J
Poznanović
Goran
G
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D003432:Cross-Linking Reagents; D004247:DNA; D047468:Immunoprecipitation; D034904:Lamin Type A / Q000378:metabolism*; D034921:Lamin Type B / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D008297:Male; D015530:Nuclear Matrix / Q000378:metabolism; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D025941:Protein Interaction Mapping; D051381:Rats; D017208:Rats, Wistar; D013347:Subcellular Fractions
D003432:Cross-Linking Reagents; D034904:Lamin Type A; D034921:Lamin Type B; C440308:lamin C; D004247:DNA; C499322:Parp1 protein, rat; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases