Nuclear relocalization of the pre-mRNA splicing factor PSF during apoptosis involves hyperphosphorylation, masking of antigenic epitopes, and changes in protein interactions.
The spatial nuclear organization of regulatory proteins often reflects their functional state. PSF, a factor essential for pre-mRNA splicing, is visualized by the B92 mAb as discrete nuclear foci, which disappeared during apoptosis. Because this mode of cell death entails protein degradation, it was considered that PSF, which like other splicing factors is sensitive to proteolysis, might be degraded. Nonetheless, during the apoptotic process, PSF remained intact and was N-terminally hyperphosphorylated on serine and threonine residues. Retarded gel migration profiles suggested differential phosphorylation of the molecule in mitosis vs. apoptosis and under-phosphorylation during blockage of cells at G1/S. Experiments with the use of recombinant GFP-tagged PSF provided evidence that in the course of apoptosis the antigenic epitopes of PSF are masked and that PSF reorganizes into globular nuclear structures. In apoptotic cells, PSF dissociated from PTB and bound new partners, including the U1--70K and SR proteins and therefore may acquire new functions.
Shav-Tal
Y
Y
Department of Molecular Cell Biology, The Weizmann Institute of Science, Rehovot, Israel.
Cohen
M
M
Lapter
S
S
Dye
B
B
Patton
J G
JG
Vandekerckhove
J
J
Zipori
D
D
D021581:Active Transport, Cell Nucleus / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001483:Base Sequence; D001854:Bone Marrow Cells / Q000378:metabolism; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology*; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003609:Dactinomycin / Q000494:pharmacology; D005260:Female; D017930:Genes, Reporter; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D000072670:PTB-Associated Splicing Factor; D010766:Phosphorylation; D012326:RNA Splicing / Q000502:physiology*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016415:Sequence Alignment
D019384:Nucleic Acid Synthesis Inhibitors; D000072670:PTB-Associated Splicing Factor; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D003609:Dactinomycin