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PMID:9348540

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Citation

Grandi, P, Dang, T, Pané, N, Shevchenko, A, Mann, M, Forbes, D and Hurt, E (1997) Nup93, a vertebrate homologue of yeast Nic96p, forms a complex with a novel 205-kDa protein and is required for correct nuclear pore assembly. Mol. Biol. Cell 8:2017-38

Abstract

Yeast and vertebrate nuclear pores display significant morphological similarity by electron microscopy, but sequence similarity between the respective proteins has been more difficult to observe. Herein we have identified a vertebrate nucleoporin, Nup93, in both human and Xenopus that has proved to be an evolutionarily related homologue of the yeast nucleoporin Nic96p. Polyclonal antiserum to human Nup93 detects corresponding proteins in human, rat, and Xenopus cells. Immunofluorescence and immunoelectron microscopy localize vertebrate Nup93 at the nuclear basket and at or near the nuclear entry to the gated channel of the pore. Immunoprecipitation from both mammalian and Xenopus cell extracts indicates that a small fraction of Nup93 physically interacts with the nucleoporin p62, just as yeast Nic96p interacts with the yeast p62 homologue. However, a large fraction of vertebrate Nup93 is extracted from pores and is also present in Xenopus egg extracts in complex with a newly discovered 205-kDa protein. Mass spectrometric sequencing of the human 205-kDa protein reveals that this protein is encoded by an open reading frame, KIAAO225, present in the human database. The putative human nucleoporin of 205 kDa has related sequence homologues in Caenorhabditis elegans and Saccharomyces cerevisiae. The analyze the role of the Nup93 complex in the pore, nuclei were assembled that lack the Nup93 complex after immunodepletion of a Xenopus nuclear reconstitution extract. The Nup93-complex-depleted nuclei are clearly defective for correct nuclear pore assembly. From these experiments, we conclude that the vertebrate and yeast pore have significant homology in their functionally important cores and that, with the identification of Nup93 and the 205-kDa protein, we have extended the knowledge of the nearest-neighbor interactions of this core in both yeast and vertebrates.

Links

PubMed PMC25664

Keywords

Amino Acid Sequence; Animals; Biological Evolution; Cell Nucleus/chemistry; Cell Nucleus/metabolism; Conserved Sequence; Fluorescent Antibody Technique, Indirect; Fungal Proteins; Humans; Ion Channels/chemistry; Membrane Proteins; Molecular Sequence Data; Nuclear Envelope/chemistry; Nuclear Envelope/metabolism; Nuclear Pore Complex Proteins; Nuclear Proteins/genetics; Nuclear Proteins/metabolism; Saccharomyces cerevisiae/chemistry; Saccharomyces cerevisiae Proteins; Sequence Homology, Amino Acid; Staining and Labeling; Vertebrates/genetics; Xenopus; Xenopus Proteins

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:NUP98

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q92621

F

Seeded From UniProt

complete

HUMAN:NU205

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P52948

F

Seeded From UniProt

complete

HUMAN:NU205

involved_in

GO:0006913: nucleocytoplasmic transport

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

HUMAN:NU205

part_of

GO:0005643: nuclear pore

ECO:0000303: author statement without traceable support used in manual assertion

C

Seeded From UniProt

complete

HUMAN:NUP98

involved_in

GO:0006913: nucleocytoplasmic transport

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

HUMAN:NUP98

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000303: author statement without traceable support used in manual assertion

F

Seeded From UniProt

complete

HUMAN:NUP98

involved_in

GO:0006999: nuclear pore organization

ECO:0000303: author statement without traceable support used in manual assertion

P

Seeded From UniProt

complete

HUMAN:NUP98

involved_in

GO:0006260: DNA replication

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:NUP98

part_of

GO:0005643: nuclear pore

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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