Association of nucleoplasmin with transcription products as revealed by immunolocalization in the amphibian oocyte.

Abstract

The oocyte nucleus of Pleurodeles waltlii contains a major 32,000-mol-wt acidic protein which is called nucleoplasmin. Rabbit antibodies were raised against total nuclear proteins from Pleurodeles oocytes. Affinity-purified antibodies directed against nucleoplasmin were prepared using antigens bound to nitrocellulose paper. The specificity of the antibody was controlled on two-dimensional electrophoretic gels of nuclear proteins. The intracellular distribution of nucleoplasmin was analyzed by direct immunofluoresence and the immunogold technique in light and electron microscopy. The antibody was tested (a) on a spread of the nuclear content prepared in the presence of calcium, (b) on the nuclear content spread in the presence of phalloidin so that an actin network appeared, and (c) on a spread of nuclei from oocytes previously treated by actinomycin D. In all cases, nucleoplasmin appeared to be localized on the lamp-brush loops, i.e., on the sites of transcription and, more specifically, on the ribonucleoprotein (RNP) particles; this protein was also associated with the RNP particles of the nuclear sap (free or inserted in the actin network). Nucleoplasmin was localized on large RNP particles that appeared when transcription was blocked. We never found this protein on the chromosome axis. These results suggest that nucleoplasmin may play a role in transcriptional activity.