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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...

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Autores principales: Nakamura, Masafumi, Masuda, Hirohisa, Horii, Johji, Kuma, Kei-ichi, Yokoyama, Nobuhiko, Ohba, Tomoyuki, Nishitani, Hideo, Miyata, Takashi, Tanaka, Masao, Nishimoto, Takeharu
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
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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.
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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
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