Differential nuclear localization and nuclear matrix association of the splicing factors PSF and PTB.
A monoclonal antibody raised against nuclear matrix proteins detected a protein of basic pI in human nuclear matrix protein samples of various cellular origin. The ubiquitously occurring (common) nuclear matrix protein was identified as splicing factor PSF (PTB associated splicing factor). The interaction between the splicing factors PSF and PTB/hnRNP I was confirmed by co-immunoprecipitation from nuclear salt extracts. However, the nuclear localization of PSF and PTB and their distribution in subnuclear fractions differed markedly. Isolated nuclear matrices contained the bulk of PSF, but only minor amounts of PTB. In confocal microscopy both proteins appeared in speckles, the majority of which did not co-localize. Removing a large fraction of the soluble PTB structures by salt extraction revealed some colocalization of the more stable PTB fraction with PSF. These PTB/PSF complexes as well as the observed PSF-PTB interaction may reflect the previously reported presence of PTB and PSF in spliceosomal complexes during RNA processing. The present data, however, point to different cellular distribution and nuclear matrix association of the majority of PSF and PTB.
Meissner
M
M
Institute of Tumor Biology-Cancer Research, University of Vienna, A-1090 Vienna, Austria.
Dechat
T
T
Gerner
C
C
Grimm
R
R
Foisner
R
R
Sauermann
G
G
D000911:Antibodies, Monoclonal / Q000276:immunology; D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005455:Fluorescent Antibody Technique; D018922:HL-60 Cells; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D015530:Nuclear Matrix / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D000072670:PTB-Associated Splicing Factor; D038941:Polypyrimidine Tract-Binding Protein; D011233:Precipitin Tests; D012326:RNA Splicing; D016601:RNA-Binding Proteins / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D000072670:PTB-Associated Splicing Factor; D016601:RNA-Binding Proteins; D038941:Polypyrimidine Tract-Binding Protein