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Nuclear pore complex: novel glycoprotein constituents

Thesis/Dissertation ·
OSTI ID:6292253
The author describes the production of anti-nuclear monoclonal antibodies, and their use in identifying some of the constituents of the nuclear pore complex. Studies are described using an antibody that recognizes predominantly a 62 kD protein (p62) of rat liver nuclei. This protein remains associated with the nuclear pore complex-lamina fraction that results from treatment of nuclei with DNase, RNase and nonionic detergent. Immunofluorescence microscopy revealed a strikingly punctate pattern of nuclear rim staining. Using immunogold electron microscopy, p62 was specifically localized to the pore complex. Pulse chase analysis of labelled tissue culture cells showed that p62 is synthesized as a soluble cytoplasmic precursor. Additionally, p62 is posttranslationally modified by the addition of GlcNAc residues. These proteins have been shown to contain O-linked monosaccharidic GlcNAc residues. Pulse chase labelling of tissue culture cells with /sup 35/S-methionine indicates that most of the sugar is added within 5 min of synthesis, when p62 is soluble and cytosolic. Tunicamycin does not prevent glycosylation and the M/sub r/ of p62 is not affected by endoglycosidase H. Thus, the addition of GlcNAc appears to be carried out by a cytoplasmically disposed transferase and is clearly distinct from the N- and O-linked glycosylation pathways in the endoplasmic reticulum and Golgi complex.
Research Organization:
Rockefeller Univ., New York (USA)
OSTI ID:
6292253
Country of Publication:
United States
Language:
English