Distribution of sugar-binding sites within interphase nuclei and mitotic chromosomes of a human cell line.
Using gold labelled neoglycoproteins containing either alpha-D-glucose, N-acetyl-beta-D-glucosamine, alpha-D-mannose, 6-phospho-alpha-D-mannose, and alpha-L-fucose (BSA), we investigated their intranuclear binding sites in the TG human cell line. Although gold-labelled BSA did not give any noticeable labelling, the presence of 1% free BSA in the medium containing the gold labelled neoglycoproteins was revealed to be a key factor of the labelling. During interphase in the presence of free BSA most of the labelling was detected in the nucleoplasm. The border of the condensed chromatin, known to be the site of hnRNA synthesis as well as the interchromatin areas enriched in RNPs were labelled. Condensed chromatin also contained binding-sites. The nucleolus was seen to present low labelling in comparison with the labelling observed over the nucleoplasm. These nucleolar binding sites were located both in the dense fibrillar and granular components. No labelling could be detected over the fibrillar centers which are very conspicuous in this cell line. During mitosis sugar-binding sites were observed over the chromosomes. Data reported here show for the first time that lectin-like proteins and chromatin components are colocalized both during interphase and mitosis. In addition, within the nucleolus the presence of sugar-binding proteins was seen to be restricted to the dense fibrillar and granular components.
Bourgeois
C A
CA
Départment de Biochimie des Glycoconjugués, Centre de Biophysique Moléculaire, Orléans, France.
Monsigny
M
M
D050260:Carbohydrate Metabolism; D002352:Carrier Proteins / Q000032:analysis*; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry* / Q000648:ultrastructure; D002875:Chromosomes / Q000737:chemistry* / Q000648:ultrastructure; D006023:Glycoproteins / Q000378:metabolism; D006046:Gold; D006801:Humans; D007399:Interphase / Q000502:physiology*; D008938:Mitosis / Q000502:physiology*; D009697:Nucleolus Organizer Region / Q000737:chemistry / Q000648:ultrastructure; D011956:Receptors, Cell Surface
D002352:Carrier Proteins; D006023:Glycoproteins; D011956:Receptors, Cell Surface; C055213:saccharide-binding proteins; D006046:Gold