Coilin shuttles between the nucleus and cytoplasm in Xenopus oocytes.
Coiled bodies are discrete nuclear organelles often identified by the marker protein p80-coilin. Because coilin is not detected in the cytoplasm by immunofluorescence and Western blotting, it has been considered an exclusively nuclear protein. In the Xenopus germinal vesicle (GV), most coilin actually resides in the nucleoplasm, although it is highly concentrated in 50-100 coiled bodies. When affinity-purified anti-coilin antibodies were injected into the cytoplasm of oocytes, they could be detected in coiled bodies within 2-3 h. Coiled bodies were intensely labeled after 18 h, whereas other nuclear organelles remained negative. Because the nuclear envelope does not allow passive diffusion of immunoglobulins, this observation suggests that anti-coilin antibodies are imported into the nucleus as an antigen-antibody complex with coilin. Newly synthesized coilin is not required, because cycloheximide had no effect on nuclear import and subsequent targeting of the antibodies. Additional experiments with myc-tagged coilin and myc-tagged pyruvate kinase confirmed that coilin is a shuttling protein. The shuttling of Nopp140, NO38/B23, and nucleolin was easily demonstrated by the targeting of their respective antibodies to the nucleoli, whereas anti-SC35 did not enter the germinal vesicle. We suggest that coilin, perhaps in association with Nopp140, may function as part of a transport system between the cytoplasm and the coiled bodies.
Bellini
M
M
Department of Embryology, Carnegie Institution, Baltimore, Maryland 21210, USA.
Gall
J G
JG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000936:Antigen-Antibody Complex / Q000378:metabolism; D001692:Biological Transport; D002467:Cell Nucleus / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D008845:Microinjections; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism*; D000090243:Nucleophosmin; D009865:Oocytes / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D014981:Xenopus / Q000378:metabolism*; D000097796:Nucleolin
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D003513:Cycloheximide; D009687:Nuclear Proteins; D000090243:Nucleophosmin; D010750:Phosphoproteins; D016271:Proto-Oncogene Proteins c-myc; D016601:RNA-Binding Proteins; D000097796:Nucleolin; C075564:NOLC1 protein, human; C068921:p80-coilin