RRC ID 48894
Author Konishi HA, Asai S, Watanabe TM, Yoshimura SH.
Title In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis.
Journal Sci Rep
Abstract The central channel of the nuclear pore complex (NPC) is occupied by non-structured polypeptides with a high content of Phe-Gly (FG) motifs. This protein-rich environment functions as an entropic barrier that prevents the passage of molecules, as well as the binding sites for karyopherins, to regulate macromolecular traffic between the nucleoplasm and the cytoplasm. In this study, we expressed individual Nups fused with a crowding-sensitive probe (GimRET) to determine the spatial distribution of protein-rich domains within the central channel in vivo, and characterize the properties of the entropic barrier. Analyses of the probe signal revealed that the central channel contains two protein-rich domains at both the nucleoplasmic and cytoplasmic peripheries, and a less-crowded central cavity. Karyopherins and other soluble proteins are not the constituents of the protein-rich domains. The time-lapse observation of the post-mitotic reassembly process also revealed how individual protein-rich domains are constructed by a sequential assembly of nucleoporins.
Volume 7(1)
Pages 5709
Published 2017-7-18
DOI 10.1038/s41598-017-05959-w
PII 10.1038/s41598-017-05959-w
PMID 28720791
PMC PMC5515885
MeSH Active Transport, Cell Nucleus / physiology* Binding Sites Cell Nucleus / metabolism* Cytoplasm / metabolism Escherichia coli / metabolism HeLa Cells Humans Karyopherins / metabolism Macromolecular Substances Nuclear Pore / physiology* Nuclear Pore Complex Proteins / metabolism*
IF 3.998
Times Cited 4
DNA material pPAL7_GimRET (RDB14203) pGimRET (RDB14204).