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Co‐localization of PARP‐1 and lamin B in the nuclear architecture: A halo‐fluorescence‐ and confocal‐microscopy study
Author(s) -
Vidaković Melita,
Koester Mario,
Goetze Sandra,
Winkelmann Silke,
Klar Martin,
Poznanović Goran,
Bode Juergen
Publication year - 2005
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.20516
Subject(s) - lamin , nuclear lamina , confocal microscopy , poly adp ribose polymerase , fluorescence microscope , nucleoplasm , nucleolus , confocal , microbiology and biotechnology , biology , biophysics , cell nucleus , microscopy , apoptosis , fluorescence , chemistry , dna , polymerase , biochemistry , nuclear protein , nucleus , pathology , transcription factor , gene , physics , geometry , mathematics , quantum mechanics , medicine
A functional interaction between poly(ADP‐ribose) polymerase‐1 (PARP‐1) and lamin B has recently been proposed by nuclear fractionation, crosslinking, and immunoprecipitation experiments. Here we use fluorescence microscopy to verify and extend these findings. We analyze nuclear halo preparations by fluorescence in situ immuno staining (FISIS), which shares attributes with traditional nuclear fractionation techniques, and by confocal laser scanning microscopy (CLSM). The results agree in that a major part of the enzyme co‐localizes with lamin B under physiological conditions, where PARP‐1 only has basal activity. After DNA damage and the associated activation of PARP‐1, and during the subsequent entry into apoptosis, dramatic changes occur: a gradual release of the enzyme from the lamina, accompanied by its accumulation in nucleoli. Our observations are in line with biochemical evidence for lamin B‐PARP‐1 interactions under physiological conditions and suggest ways by which these interactions are modified to support PARP‐functions in damage and its fate in apoptosis. © 2005 Wiley‐Liss, Inc.