Rekonstitution des symmetrischen Gerüstes sowie Strukturaufklärung der wichtigsten Subkomplexe des Kernporen Komplexes
Zusammenfassung der Projektergebnisse
The nuclear pore complex (NPC) serves as the sole gateway for bidirectional nucleocytoplasmic transport in eukaryotic cells. Despite half a century of extensive structural characterization, the atomic architecture of the NPC remained unknown. This project aimed at the biochemical dissection and reconstitution of the symmetric core of the NPC as well as the structural elucidation of its main sub-complexes. A major breakthrough in the structural elucidation of the NPC coat-forming coat nucleoporin complex (CNC) was made describing the X-ray structure of a reconstituted hexameric CNC from S.cerevisiae that could readily be docked into a published Cryo- Eletron tomography map of the human NPC. The placement of the CNC structure yielded a model that explains ~16 MDa of the mass of the NPC and elucidated the architecture of its membrane coat. Furthermore, the project let to the structural elucidation of 11 different structures of the second largest adapter nucleoporin complex found at the core of the NPC. A reconstitution of both the CNC and adapter nucleoporin complex resulted in an in vitro assembly system for a 1.6 MDa protomer of the entire symmetric core of the NPC explaining the molecular details of one of natures biggest transport organelle. The work presents one of a view examples of a reconstituted protein assembly of this complexity that was build up from individually expressed and purified proteins. Parts of the work were featured as a science highlight at Caltech and the advanced photon source in Chicago: http://www.caltech.edu/news/caltech-biochemist-sheds-light-structure-key-cellulargatekeeper-45681 https://www1.aps.anl.gov/APS-Science-Highlight/2015/A-Key-Cellular-Gatekeeper
Projektbezogene Publikationen (Auswahl)
- (2014). Evidence for an evolutionary relationship between the large adaptor nucleoporin Nup192 and karyopherins. PNAS. 111, 2530-2535
Stuwe, T., Lin, DH., Collins, L.N., Hurt, E., Hoelz, A.
(Siehe online unter https://doi.org/10.1073/pnas.1311081111) - (2015). Architecture of the nuclear pore complex coat. Science. 347, 1148-1152
Stuwe, T., Correia, A.R., Lin, DH., Collins, L.N., Paduch, M., Lu, V.T., Kosiakoff, A.A., Hoelz, A.
(Siehe online unter https://doi.org/10.1126/science.aaa4136)
