title: Structure-function mapping of a heptameric module in the nuclear pore complex.
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.
authors.affiliation: Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Javier
authors.initials: J
authors.lastname: Fernandez-Martinez
authors.forename: Jeremy
authors.initials: J
authors.lastname: Phillips
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Sekedat
authors.forename: Ruben
authors.initials: R
authors.lastname: Diaz-Avalos
authors.forename: Javier
authors.initials: J
authors.lastname: Velazquez-Muriel
authors.forename: Josef D
authors.initials: JD
authors.lastname: Franke
authors.forename: Rosemary
authors.initials: R
authors.lastname: Williams
authors.forename: David L
authors.initials: DL
authors.lastname: Stokes
authors.forename: Brian T
authors.initials: BT
authors.lastname: Chait
authors.forename: Andrej
authors.initials: A
authors.lastname: Sali
authors.forename: Michael P
authors.initials: MP
authors.lastname: Rout
mesh_terms: D008958:Models, Molecular; D022022:Nuclear Pore / Q000502:physiology* / Q000648:ultrastructure; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D013329:Structure-Activity Relationship
chemical_list: C097894:NUP84 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D029701:Saccharomyces cerevisiae Proteins