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PMID:22331846

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Citation

Fernandez-Martinez, J, Phillips, J, Sekedat, MD, Diaz-Avalos, R, Velazquez-Muriel, J, Franke, JD, Williams, R, Stokes, DL, Chait, BT, Sali, A and Rout, MP (2012) Structure-function mapping of a heptameric module in the nuclear pore complex. J. Cell Biol. 196:419-34

Abstract

The nuclear pore complex (NPC) is a multiprotein assembly that serves as the sole mediator of nucleocytoplasmic exchange in eukaryotic cells. In this paper, we use an integrative approach to determine the structure of an essential component of the yeast NPC, the ~600-kD heptameric Nup84 complex, to a precision of ~1.5 nm. The configuration of the subunit structures was determined by satisfaction of spatial restraints derived from a diverse set of negative-stain electron microscopy and protein domain-mapping data. Phenotypic data were mapped onto the complex, allowing us to identify regions that stabilize the NPC's interaction with the nuclear envelope membrane and connect the complex to the rest of the NPC. Our data allow us to suggest how the Nup84 complex is assembled into the NPC and propose a scenario for the evolution of the Nup84 complex through a series of gene duplication and loss events. This work demonstrates that integrative approaches based on low-resolution data of sufficient quality can generate functionally informative structures at intermediate resolution.

Links

PubMed PMC3283990 Online version:10.1083/jcb.201109008

Keywords

Models, Molecular; Nuclear Pore/physiology; Nuclear Pore/ultrastructure; Nuclear Pore Complex Proteins/chemistry; Nuclear Pore Complex Proteins/genetics; Nuclear Pore Complex Proteins/metabolism; Protein Conformation; Protein Structure, Tertiary; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/growth & development; Saccharomyces cerevisiae/metabolism; Saccharomyces cerevisiae Proteins/chemistry; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae Proteins/metabolism; Sequence Deletion; Structure-Activity Relationship

Significance

Annotations

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

YEAST:NU120

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

YEAST:NU120

involved_in

GO:0051664: nuclear pore localization

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:NU133

involved_in

GO:0051664: nuclear pore localization

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:NUP85

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

YEAST:NU145

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

YEAST:NUP84

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

YEAST:NU133

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

YEAST:NU133

involved_in

GO:0031081: nuclear pore distribution

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

See also

References

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