Bleyet al., Science 376 , eabm9129 (2022) 10 June 2022 4of18
Fig. 2. Conserved modular architecture and RNA-binding properties of the
human CF nups.(A) Cross-sectional schematic of the human NPC architecture.
(B) Domain structures of human CF nups. Nomenclature ofH. sapiensand
C. thermophilumnup orthologs is indicated. (C) Biochemical reconstitution of the
~310-kDa heterohexameric human CFNC. SEC-MALS interaction analysis of
NUP88•NUP214•NUP62 (blue), DDX19 (ADP) (red), RAE1•NUP98 (cyan), and
their preincubation (green). Measured molecular masses are indicated, with
theoretical masses in parentheses. SDS-PAGE gel strips of peak fractions
visualized by Coomassie brilliant blue staining are shown. (D) Summary of
pairwise SEC-MALS interaction analyses between human CF nups. (E) Schematic
summary of the human CFNC architecture and the CF nup interaction
network. (F) Human CF nup domains and complexes were assayed for
binding to ssRNA and dsDNA probes by EMSA. Input proteins resolved by
SDS-PAGE were visualized by Coomassie brilliant blue staining. Qualitative
assessment of nucleic acid binding is denoted by color-coded boxes. (G and
H) EMSAs with ssRNA titrated against (G) metazoan-specific NUP358NTD
and NUP358RanBD-IV•Ran(GMPPNP), and (H) the indicatedH. sapiensCFNC
subcomplexes and theirC. thermophilumorthologs.RESEARCH | STRUCTURE OF THE NUCLEAR PORE