Cargando…
When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to γ-Tubulin
A novel human protein with a molecular mass of 55 kD, designated RanBPM, was isolated with the two-hybrid method using Ran as a bait. Mouse and hamster RanBPM possessed a polypeptide identical to the human one. Furthermore, Saccharomyces cerevisiae was found to have a gene, YGL227w, the COOH-termina...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
1998
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2132962/ https://www.ncbi.nlm.nih.gov/pubmed/9817760 |
_version_ | 1782142563246931968 |
---|---|
author | Nakamura, Masafumi Masuda, Hirohisa Horii, Johji Kuma, Kei-ichi Yokoyama, Nobuhiko Ohba, Tomoyuki Nishitani, Hideo Miyata, Takashi Tanaka, Masao Nishimoto, Takeharu |
author_facet | Nakamura, Masafumi Masuda, Hirohisa Horii, Johji Kuma, Kei-ichi Yokoyama, Nobuhiko Ohba, Tomoyuki Nishitani, Hideo Miyata, Takashi Tanaka, Masao Nishimoto, Takeharu |
author_sort | Nakamura, Masafumi |
collection | PubMed |
description | A novel human protein with a molecular mass of 55 kD, designated RanBPM, was isolated with the two-hybrid method using Ran as a bait. Mouse and hamster RanBPM possessed a polypeptide identical to the human one. Furthermore, Saccharomyces cerevisiae was found to have a gene, YGL227w, the COOH-terminal half of which is 30% identical to RanBPM. Anti-RanBPM antibodies revealed that RanBPM was localized within the centrosome throughout the cell cycle. Overexpression of RanBPM produced multiple spots which were colocalized with γ-tubulin and acted as ectopic microtubule nucleation sites, resulting in a reorganization of microtubule network. RanBPM cosedimented with the centrosomal fractions by sucrose- density gradient centrifugation. The formation of microtubule asters was inhibited not only by anti- RanBPM antibodies, but also by nonhydrolyzable GTP-Ran. Indeed, RanBPM specifically interacted with GTP-Ran in two-hybrid assay. The central part of asters stained by anti-RanBPM antibodies or by the mAb to γ-tubulin was faded by the addition of GTPγS-Ran, but not by the addition of anti-RanBPM anti- bodies. These results provide evidence that the Ran-binding protein, RanBPM, is involved in microtubule nucleation, thereby suggesting that Ran regulates the centrosome through RanBPM. |
format | Text |
id | pubmed-2132962 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1998 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21329622008-05-01 When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to γ-Tubulin Nakamura, Masafumi Masuda, Hirohisa Horii, Johji Kuma, Kei-ichi Yokoyama, Nobuhiko Ohba, Tomoyuki Nishitani, Hideo Miyata, Takashi Tanaka, Masao Nishimoto, Takeharu J Cell Biol Regular Articles A novel human protein with a molecular mass of 55 kD, designated RanBPM, was isolated with the two-hybrid method using Ran as a bait. Mouse and hamster RanBPM possessed a polypeptide identical to the human one. Furthermore, Saccharomyces cerevisiae was found to have a gene, YGL227w, the COOH-terminal half of which is 30% identical to RanBPM. Anti-RanBPM antibodies revealed that RanBPM was localized within the centrosome throughout the cell cycle. Overexpression of RanBPM produced multiple spots which were colocalized with γ-tubulin and acted as ectopic microtubule nucleation sites, resulting in a reorganization of microtubule network. RanBPM cosedimented with the centrosomal fractions by sucrose- density gradient centrifugation. The formation of microtubule asters was inhibited not only by anti- RanBPM antibodies, but also by nonhydrolyzable GTP-Ran. Indeed, RanBPM specifically interacted with GTP-Ran in two-hybrid assay. The central part of asters stained by anti-RanBPM antibodies or by the mAb to γ-tubulin was faded by the addition of GTPγS-Ran, but not by the addition of anti-RanBPM anti- bodies. These results provide evidence that the Ran-binding protein, RanBPM, is involved in microtubule nucleation, thereby suggesting that Ran regulates the centrosome through RanBPM. The Rockefeller University Press 1998-11-16 /pmc/articles/PMC2132962/ /pubmed/9817760 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Regular Articles Nakamura, Masafumi Masuda, Hirohisa Horii, Johji Kuma, Kei-ichi Yokoyama, Nobuhiko Ohba, Tomoyuki Nishitani, Hideo Miyata, Takashi Tanaka, Masao Nishimoto, Takeharu When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to γ-Tubulin |
title | When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to γ-Tubulin |
title_full | When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to γ-Tubulin |
title_fullStr | When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to γ-Tubulin |
title_full_unstemmed | When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to γ-Tubulin |
title_short | When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to γ-Tubulin |
title_sort | when overexpressed, a novel centrosomal protein, ranbpm, causes ectopic microtubule nucleation similar to γ-tubulin |
topic | Regular Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2132962/ https://www.ncbi.nlm.nih.gov/pubmed/9817760 |
work_keys_str_mv | AT nakamuramasafumi whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin AT masudahirohisa whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin AT horiijohji whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin AT kumakeiichi whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin AT yokoyamanobuhiko whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin AT ohbatomoyuki whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin AT nishitanihideo whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin AT miyatatakashi whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin AT tanakamasao whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin AT nishimototakeharu whenoverexpressedanovelcentrosomalproteinranbpmcausesectopicmicrotubulenucleationsimilartogtubulin |