<> "The repository administrator has not yet configured an RDF license."^^ . <> . . "In cellulo architecture of the nuclear pore complex"^^ . "Nuclear pore complexes (NPCs) reside at the nuclear envelope (NE) where they mediate\r\nnucleocytoplasmic exchange. They are large macromolecular assemblies built by several\r\nhundred protein building blocks, so-called nucleoporins (Nups), which form numerous\r\nsubcomplexes to fuse the inner and outer nuclear membranes and form massive 8-fold\r\nrotational symmetric nuclear pore. Solving the structure of NPCs as a whole imposes a\r\ntremendous challenge. Yet, integrated structural analysis approaches led to the determination of the human, S. cerevisiae (sc) and the green algae C. reinhardtii NPC scaffold\r\narchitecture, in situ, in vitro and in cellulo, respectively. It is apparent that NPCs across\r\nspecies, although varying in their subcomplex composition and stoichiometry, follow a\r\ncommon architectural building concept. Three distinct rings stacked across the NE form\r\nthe NPC scaffold. The inner ring forms a central channel, and together with the flanking,\r\ncytoplasmic and nuclear rings, ensures the nuclear pore stability. The outer rings are built\r\nby a head-to-tail arrangement of the so-called Y-complex. In humans, two concentric rings\r\nform each of the two outer rings while in S. cerevsiaie only one Y-complex ring on each\r\nside of the NPC is found. In algae, two Y-rings form the nuclear ring while only one ring is\r\npresent on the cytoplasmic side. The metazoan-specific Nup358 mediates the cytoplasmic\r\nY-complex dimerization in human, however, the corresponding factor mediating the nuclear\r\nring dimerization remains elusive in all species to date. It is suspected that NPCs might\r\nadapt their central channel diameter in response to a large variety of cues to alter nuclear\r\ntransport. However, what exactly causes such a conformational change within actively\r\ntransporting cells and how it is mediated on a molecular level is not known.\r\nHere, I solved the structure of the S. pombe (sp) NPC within intact cells. I show\r\nhow a split Y-complex interface on the cytoplasmic face of the spNPC breaks the outer\r\nring conformation and thus challenges the long-standing dogma of a three-ringed NPC\r\narchitecture. I found two concentric Y-rings on the nuclear side of the spNPC. A candidate\r\nprotein to mediate this interface was the nucleoporin Ely5, which is only found in species\r\nshowing two concentric nuclear rings. However, upon analyzing the NPC architecture of an\r\nely5Δ knockout strain, I could rule out Ely5 as a sufficient Y-complex dimerization factor.\r\nIn collaboration with Matteo Allegretti, we solved an in cellulo scNPC structure and show\r\nhow it differs from the previously proposed in vitro derived architecture. We re-orient\r\nthe previously proposed mRNA export platform model anchored at the cytoplasmic ring\r\n9\r\nto be more in line with the spatiotemporal events coordinating mRNP nuclear export. I\r\nfurther present an in-depth comparison between the two yeast to the human and algal\r\nNPC architectures revealing that while the inner ring remains highly conserved, the\r\narchitecture of the outer rings, the mRNA export platform, as well as the connections\r\nbetween subcomplexes varies more than previously anticipated. To understand how NPCs\r\nadopt their conformation during active nuclear transport on a molecular level, I solved\r\na spNPC structure under energy depletion conditions where active nuclear transport is\r\nthought to be completely halted. This analysis exhibits a constricted NPC conformation,\r\nrevealing how the NPC scaffold undergoes a conformational change on a molecular level in\r\nresponse to a well-defined cue. Finally, I investigate how the transmembrane nucleoporin\r\nPom152 (hsGP210), a protein known to form a ring surrounding the NPC within the NE\r\nlumen, might be involved in NPC diameter control. By analyzing a human gp210Δ cell\r\nline I could show that while GP210 is dispensable for proper NPC biogenesis, the missing\r\nluminal ring did not influence the human NPC diameter observed under the conditions\r\ndescribed here."^^ . "2020" . . . . . . . "Christian Eugen"^^ . "Zimmerli"^^ . "Christian Eugen Zimmerli"^^ . . . . . . "In cellulo architecture of the nuclear pore complex (PDF)"^^ . . . "PhD_Thesis_Christian_Zimmerli.pdf"^^ . . . "In cellulo architecture of the nuclear pore complex (Other)"^^ . . . . . . "indexcodes.txt"^^ . . . "In cellulo architecture of the nuclear pore complex (Other)"^^ . . . . . . "lightbox.jpg"^^ . . . "In cellulo architecture of the nuclear pore complex (Other)"^^ . . . . . . "preview.jpg"^^ . . . "In cellulo architecture of the nuclear pore complex (Other)"^^ . . . . . . "medium.jpg"^^ . . . "In cellulo architecture of the nuclear pore complex (Other)"^^ . . . . . . "small.jpg"^^ . . "HTML Summary of #28356 \n\nIn cellulo architecture of the nuclear pore complex\n\n" . "text/html" . . . "570 Biowissenschaften, Biologie"@de . "570 Life sciences"@en . .