Nucleoplasmin binds to nuclear pore filaments and accumulates in specific regions of the nucleolar cortex.
Nucleoplasmin is a karyophilic protein that is involved in nucleosome formation and decondensation of chromatin, although other precise functions and modes of action of this molecule are still poorly understood. In the present paper we describe a novel nucleocytoplasmic transport assay that has enabled us to study the nuclear distribution of nucleoplasmin following its transport into the nucleus. Single Xenopus laevis oocyte nuclei were isolated and incubated with Xenopus egg extract containing colloidal gold-conjugated nucleoplasmin. After a period of incubation, each individual nucleus was processed for electron microscopy. The nuclear accumulation of nucleoplasmin was dependent upon the karyophilic properties of the protein, since BSA-conjugated gold particles did not enter the nuclear interior under the same experimental conditions. Once inside the nucleus, nucleoplasmin was detected in tracks emanating from the nuclear pores and reaching the nucleolus. Additionally, we found a striking accumulation of nucleoplasmin in specific areas of the nucleolar cortex. These perinucleolar regions were surrounded by areas of electron density similar to that of the fibrillar centers. Our results indicate that nucleoplasmin may play an important role in the transcription of ribosomal precursors. Moreover, this nucleocytoplasmic transport assay will enable the determination of the precise intranuclear localization of other karyophilic proteins.
Andrade
R
R
Department of Cell Biology and General Service of Analytical and High Resolution Microscopy, University of the Basque Country, Medical School, Leioa, Vizcaya, Spain.
Arlucea
J
J
Alonso
R
R
Aréchaga
J
J
D000818:Animals; D002466:Cell Nucleolus / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D022022:Nuclear Pore / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D056489:Nucleoplasmins; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding; D014982:Xenopus laevis
D009687:Nuclear Proteins; D056489:Nucleoplasmins; D010750:Phosphoproteins