title: Structural determinants and mechanism of mammalian CRM1 allostery.
abstract: Proteins carrying nuclear export signals cooperatively assemble with the export factor CRM1 and the effector protein RanGTP. In lower eukaryotes, this cooperativity is coupled to CRM1 conformational changes; however, it is unknown if mammalian CRM1 maintains its compact conformation or shows similar structural flexibility. Here, combinations of small-angle X-ray solution scattering and electron microscopy experiments with molecular dynamics simulations reveal pronounced conformational flexibility in mammalian CRM1 and demonstrate that RanGTP binding induces association of its N- and C-terminal regions to form a toroid structure. The CRM1 toroid is stabilized mainly by local interactions between the terminal regions, rather than by global strain. The CRM1 acidic loop is key in transmitting the effect of this RanGTP-induced global conformational change to the NES-binding cleft by shifting its population to the open state, which displays enhanced cargo affinity. Cooperative CRM1 export complex assembly thus constitutes a highly dynamic process, encompassing an intricate interplay of global and local structural changes.
authors.affiliation: Abteilung für Theoretische und Computergestützte Biophysik, Max-Planck-Institut für Biophysikalische Chemie, Am Faßberg 11, 37077 Göttingen, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Dölker
authors.forename: Clement E
authors.initials: CE
authors.lastname: Blanchet
authors.forename: Béla
authors.initials: B
authors.lastname: Voß
authors.forename: David
authors.initials: D
authors.lastname: Haselbach
authors.forename: Christian
authors.initials: C
authors.lastname: Kappel
authors.forename: Thomas
authors.initials: T
authors.lastname: Monecke
authors.forename: Dmitri I
authors.initials: DI
authors.lastname: Svergun
authors.forename: Holger
authors.initials: H
authors.lastname: Stark
authors.forename: Ralf
authors.initials: R
authors.lastname: Ficner
authors.forename: Ulrich
authors.initials: U
authors.lastname: Zachariae
authors.forename: Helmut
authors.initials: H
authors.lastname: Grubmüller
authors.forename: Achim
authors.initials: A
authors.lastname: Dickmanns
mesh_terms: D000494:Allosteric Regulation; D000495:Allosteric Site; D019943:Amino Acid Substitution; D000818:Animals; D006160:Guanosine Triphosphate / Q000737:chemistry; D006801:Humans; D028884:Karyopherins / Q000737:chemistry* / Q000235:genetics; D051379:Mice; D008854:Microscopy, Electron; D056004:Molecular Dynamics Simulation; D009687:Nuclear Proteins / Q000737:chemistry; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D017433:Protein Structure, Secondary; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry* / Q000235:genetics; D053838:Scattering, Small Angle; D012996:Solutions; D014961:X-Ray Diffraction; D020931:ran GTP-Binding Protein / Q000737:chemistry; D000097606:Exportin 1 Protein
chemical_list: D006160:Guanosine Triphosphate; D028884:Karyopherins; D009687:Nuclear Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D012996:Solutions; D020931:ran GTP-Binding Protein; D000097606:Exportin 1 Protein; C120587:RAN protein, human; C091775:ran-binding protein 1