Vol. 12, Issue 12, 3904-3918, December 2001

A Novel Karyoskeletal Protein: Characterization of Protein NO145, the Major Component of Nucleolar Cortical Skeleton in Xenopus Oocytes

Sandra Kneissel,^* Werner W. Franke,^* Joseph G. Gall, Hans Heid,^* Sonja Reidenbach,^* Martina Schnölzer, Herbert Spring,^§ Hanswalter Zentgraf, and Marion S. Schmidt-Zachmann^*¶

 ^*Division of Cell Biology,  Protein Analysis Facility,  ^§Biomedical Structure Analysis Group,  Applied Tumor Virology Program, German Cancer Research Center, D-69120 Heidelberg, Germany; and  Department of Embryology, Carnegie Institution of Washington, Baltimore, Maryland 21210

The nucleolus is a ubiquitous, mostly spheroidal nuclear structure of all protein-synthesizing cells, with a well-defined functional compartmentalization. Although a number of nonribosomal proteins involved in ribosome formation have been identified, the elements responsible for the shape and internal architecture of nucleoli are still largely unknown. Here, we report the molecular characterization of a novel protein, NO145, which is a major and specific component of a nucleolar cortical skeleton resistant to high salt buffers. The amino acid sequence of this polypeptide with a SDS-PAGE mobility corresponding to M_r 145,000 has been deduced from a cDNA clone isolated from a Xenopus laevis ovary expression library and defines a polypeptide of 977 amino acids with a calculated mass of 111 kDa, with partial sequence homology to a synaptonemal complex protein, SCP2. Antibodies specific for this protein have allowed its recognition in immunoblots of karyoskeleton-containing fractions of oocytes from different Xenopus species and have revealed its presence in all stages of oogenesis, followed by a specific and rapid degradation during egg formation. Immunolocalization studies at the light and electron microscopic level have shown that protein NO145 is exclusively located in a cage-like cortical structure around the entire nucleolus, consisting of a meshwork of patches and filaments that dissociates upon reduction of divalent cations. We propose that protein NO145 contributes to the assembly of a karyoskeletal structure specific for the nucleolar cortex of the extrachromosomal nucleoli of Xenopus oocytes, and we discuss the possibility that a similar structure is present in other cells and species.