Mapping the native organization of the yeast nuclear pore complex using nuclear radial intensity measurements.
Proc Natl Acad Sci U S A
; 116(29): 14606-14613, 2019 07 16.
Article
in En
| MEDLINE
| ID: mdl-31262825
ABSTRACT
Selective transport across the nuclear envelope (NE) is mediated by the nuclear pore complex (NPC), a massive â¼100-MDa assembly composed of multiple copies of â¼30 nuclear pore proteins (Nups). Recent advances have shed light on the composition and structure of NPCs, but approaches that could map their organization in live cells are still lacking. Here, we introduce an in vivo method to perform nuclear radial intensity measurements (NuRIM) using fluorescence microscopy to determine the average position of NE-localized proteins along the nucleocytoplasmic transport axis. We apply NuRIM to study the organization of the NPC and the mobile transport machinery in budding yeast. This reveals a unique snapshot of the intact yeast NPC and identifies distinct steady-state localizations for various NE-associated proteins and nuclear transport factors. We find that the NPC architecture is robust against compositional changes and could also confirm that in contrast to Chlamydomonas reinhardtii, the scaffold Y complex is arranged symmetrically in the yeast NPC. Furthermore, NuRIM was applied to probe the orientation of intrinsically disordered FG-repeat segments, providing insight into their roles in selective NPC permeability and structure.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Saccharomyces cerevisiae
/
Nuclear Pore
/
Active Transport, Cell Nucleus
/
Saccharomyces cerevisiae Proteins
/
Intravital Microscopy
Type of study:
Prognostic_studies
Language:
En
Journal:
Proc Natl Acad Sci U S A
Year:
2019
Document type:
Article