Autoimmune serum containing an antibody against a 94 kDa nucleolar protein

doi:10.1016/0248-4900(88)90007-X

Biology of the Cell

Volume 64, Issue 3, 1988, Pages 331-341

https://doi.org/10.1016/0248-4900(88)90007-X Get rights and content

Abstract

Little information exists on how various nucleolar proteins function in ribosome biogenesis. Of special interest is that group of nucleolar proteins which are not incorporated into mature ribosomes because they are candidates for a role in the regulation of ribosome construction. Non-ribosomal nucleolar proteins can be analyzed using autoimmune sera from scleroderma patients which often contain antinuclear antibodies. One such serum, designated ScBr, is shown by indirect immunofluorescence to react specifically with nucleoli in cells of 3 different mammalian species, indicating that the antigen is at least partly conserved evolutionarily. It is not RNase-sensitive, but is completely eliminated after incubation with pronase and 2 M NaCl. Immuno-electron microscopy was carried out on Lowicryl ultrathin sections to localize the antigen. The labeling was observed over both the granular and the dense fibrillar component but not the fibrillar centers, indicating that the antigen is associated with ribosomal RNA transcription sites and ribosome assembly into precursor particles. In addition, the antibody was localized to small nucleoplasmic entities, termed dense nuclear bodies. This could indicate a relationship between nucleoli and dense nuclear bodies. By immunoprecipitation, only a single protein of 94 kDa molecular weight was revealed. By immunoblotting, the band at 94 kDa was found to be the only positive band for high ScBr dilutions. Observation of the behavior of the antigen during mitosis revealed that it became dispersed into the cytoplasm after breakdown of the nuclear envelope, lining most of the chromosomes rather that remaining associated with the NOR-chromosomes. The antigen appeared to be restored to nucleoli only in late telophase; phase-dense prenucleolar bodies of early telophase cells did not show positive staining for the antigen. During actinomycin-D RNA synthesis inhibition as well as in non-stimulated lymphocytes the positive staining is greatly decreased. These results were consistent with a role for the 94 kDa nucleolar protein in the process of preribosome assembly.

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Prenucleolar bodies (PNBs) are transitory structures which serve as building blocks for nucleoli at the transition mitosis/interphase. The assembly of PNBs and their pathway are not clearly understood. To better understand these events, the behavior of the PNB-containing PM-Scl 100 protein was compared with that of other PNB proteins. This nucleolar protein was chosen because its yeast homologue, Rrp6p exonuclease [1], is known to participate in late events in 5.8 S rRNA (ribosomal RNA) processing. There was a heterogeneous distribution of nucleolar proteins in different classes of PNBs. The PM-Scl 100 colocalized predominantly with protein B23. The PM-Scl-100-containing PNBs were translocated at later times to nucleoli as opposed to the fibrillarin-containing PNBs. Microinjections of antibodies directed against PM-Scl 100 during mitosis inhibited targeting of PM-Scl 100 to the nucleolus. However fibrillarin and protein B23 still participated in nucleolar assembly in early G1. We conclude that there are different kinds of PNBs whose translocation to the nucleoli follow ordered kinetics. Interestingly, proteins involved in late steps of processing as PM-Scl 100 are translocated late, suggesting that they are not cotranscriptionally associated with the rRNA precursors.
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Original articleAutoimmune serum containing an antibody against a 94 kDa nucleolar protein

Abstract

Exp. Cell Res.

Cell

Int. Rev. Cytol.

Exp. Cell Res.

Exp. Cell Res.

Cell

Exp. Cell Res.

Exp. Cell Res.

J. Biol. Chem.

Exp. Cell Res.

Exp. Cell Res.

J. Biol. Chem.

TIBS

Biochem. Biophys. Res. Commun.

Adv. Immunol.

Biochim. Biophys. Acta

Changes in the frequency of two types of nuclear body during the interphase of meristematic plant cells

Protoplasma

Association of antinuclear and antinucleolar antibodies in progressive systemic sclerosis

Clin. Exp. Immunol.

Maturation of a 100 kDa protein associated with preribosomes in Chinese hamster ovary cells

Mol. Biol. Rep.

Novel nucleolar antigens in autoimmune disease

J. Rheumatol.

Immunolocalization of the 100K nucleolar protein in CHO cells

Biol. Cell

Autoantibody testing: procedures and significance in systematic rheumatic diseases

Methods Achiev. Exp. Pathol.

Autoantibodies to nuclear lamin B in a patient with thrombopenia

Eur. J. Cell Biol.

Scl 70 autoantibodies from scleroderma patients recognize a 95 kDa protein identified as DNA topoisomerase I

Chromosoma

The Nucleolus and Ribosome Biogenesis

Simultaneous nucleologenesis in daughter cells during the late telophase

Biol. Cell

Ultrastructural localization of Ag-NOR stained proteins in the nucleolus during the cell cycle and in other nucleolar structures

Chromosoma

Original article
Autoimmune serum containing an antibody against a 94 kDa nucleolar protein