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.
Molecular Biology Laboratory, Institute for Biological Research, 29th November 142, 11060 Belgrade, Serbia and Montenegro, Yugoslavia.
Melita
M
Vidaković
Nevena
N
Grdović
Piera
P
Quesada
Jürgen
J
Bode
Goran
G
Poznanović
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