Actin- and protein-4.1-containing filaments link nuclear pore complexes to subnuclear organelles in Xenopus oocyte nuclei.
We imaged the interiors of relatively intact Xenopus oocyte nuclei by field emission scanning electron microscopy (feSEM) and visualized a network of filaments that attach to nuclear pore complexes and extend throughout the nucleus. Within the nucleus, these 'pore-linked filaments' (PLFs) were embedded into spherical structures 100 nm to approximately 5 microm in diameter. A subset of spheres was identified as Cajal bodies by immuno-gold labeling; the rest were inferred to be nucleoli and snurposomes both of which are abundant in Xenopus oocyte nuclei. Most PLFs were independent of chromatin. The thickness of a typical PLF was 40 nm (range, approximately 12-100 nm), including the 4 nm chromium coat. PLFs located inside the nucleus merged, bundled and forked, suggesting architectural adaptability. The PLF network collapsed upon treatment with latrunculin A, which depolymerizes actin filaments. Jasplakinolide, which stabilizes actin filaments, produced PLFs with more open substructure including individual filaments with evenly-spaced rows of radially projecting short filaments. Immuno-gold labeling of untreated oocyte nuclei showed that actin and protein 4.1 each localized on PLFs. Protein 4.1-gold epitopes were spaced at approximately 120 nm intervals along filaments, and were often paired ( approximately 70 nm apart) at filament junctions. We suggest that protein 4.1 and actin contribute to the structure of a network of heterogeneous filaments that link nuclear pore complexes to subnuclear organelles, and discuss possible functions for PLFs in nuclear assembly and intranuclear traffic.
Kiseleva
Elena
E
Department of Structural Cell Biology, Paterson Institute for Cancer Research, Christie Hospital, Manchester, M20 9BX, UK.
Drummond
Sheona P
SP
Goldberg
Martin W
MW
Rutherford
Sandra A
SA
Allen
Terence D
TD
Wilson
Katherine L
KL
D000199:Actins / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002466:Cell Nucleolus / Q000648:ultrastructure; D002467:Cell Nucleus / Q000737:chemistry* / Q000648:ultrastructure; D002843:Chromatin / Q000648:ultrastructure; D020541:Coiled Bodies / Q000378:metabolism / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000378:metabolism* / Q000648:ultrastructure; D047630:Depsipeptides / Q000494:pharmacology; D005260:Female; D007150:Immunohistochemistry; D008565:Membrane Proteins; D008855:Microscopy, Electron, Scanning; D022022:Nuclear Pore / Q000378:metabolism* / Q000648:ultrastructure; D009865:Oocytes / Q000648:ultrastructure*; D011485:Protein Binding; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines; D014981:Xenopus
D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002843:Chromatin; D003598:Cytoskeletal Proteins; D047630:Depsipeptides; D008565:Membrane Proteins; D013844:Thiazoles; D053778:Thiazolidines; C028841:erythrocyte membrane band 4.1 protein; C057531:jasplakinolide; C037067:latrunculin A