In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis

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,...

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Autores principales:	Hide A. Konishi, Suguru Asai, Tomonobu M. Watanabe, Shige H. Yoshimura
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/33865ec1dc2c427a9a46685fa3c53f6c
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spelling	oai:doaj.org-article:33865ec1dc2c427a9a46685fa3c53f6c2021-12-02T15:06:13ZIn vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis10.1038/s41598-017-05959-w2045-2322https://doaj.org/article/33865ec1dc2c427a9a46685fa3c53f6c2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05959-whttps://doaj.org/toc/2045-2322Abstract 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.Hide A. KonishiSuguru AsaiTomonobu M. WatanabeShige H. YoshimuraNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-14 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hide A. Konishi Suguru Asai Tomonobu M. Watanabe Shige H. Yoshimura In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis
description	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.
format	article
author	Hide A. Konishi Suguru Asai Tomonobu M. Watanabe Shige H. Yoshimura
author_facet	Hide A. Konishi Suguru Asai Tomonobu M. Watanabe Shige H. Yoshimura
author_sort	Hide A. Konishi
title	In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis
title_short	In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis
title_full	In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis
title_fullStr	In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis
title_full_unstemmed	In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis
title_sort	in vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/33865ec1dc2c427a9a46685fa3c53f6c
work_keys_str_mv	AT hideakonishi invivoanalysisofproteincrowdingwithinthenuclearporecomplexininterphaseandmitosis AT suguruasai invivoanalysisofproteincrowdingwithinthenuclearporecomplexininterphaseandmitosis AT tomonobumwatanabe invivoanalysisofproteincrowdingwithinthenuclearporecomplexininterphaseandmitosis AT shigehyoshimura invivoanalysisofproteincrowdingwithinthenuclearporecomplexininterphaseandmitosis
_version_	1718388503000121344

In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis

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