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Activity of Rho-family GTPases during cell division as visualized with FRET-based probes

Rho-family GTPases regulate many cellular functions. To visualize the activity of Rho-family GTPases in living cells, we developed fluorescence resonance energy transfer (FRET)–based probes for Rac1 and Cdc42 previously (Itoh, R.E., K. Kurokawa, Y. Ohba, H. Yoshizaki, N. Mochizuki, and M. Matsuda. 2...

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Autores principales: Yoshizaki, Hisayoshi, Ohba, Yusuke, Kurokawa, Kazuo, Itoh, Reina E., Nakamura, Takeshi, Mochizuki, Naoki, Nagashima, Kazuo, Matsuda, Michiyuki
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2003
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2172791/
https://www.ncbi.nlm.nih.gov/pubmed/12860967
http://dx.doi.org/10.1083/jcb.200212049
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author Yoshizaki, Hisayoshi
Ohba, Yusuke
Kurokawa, Kazuo
Itoh, Reina E.
Nakamura, Takeshi
Mochizuki, Naoki
Nagashima, Kazuo
Matsuda, Michiyuki
author_facet Yoshizaki, Hisayoshi
Ohba, Yusuke
Kurokawa, Kazuo
Itoh, Reina E.
Nakamura, Takeshi
Mochizuki, Naoki
Nagashima, Kazuo
Matsuda, Michiyuki
author_sort Yoshizaki, Hisayoshi
collection PubMed
description Rho-family GTPases regulate many cellular functions. To visualize the activity of Rho-family GTPases in living cells, we developed fluorescence resonance energy transfer (FRET)–based probes for Rac1 and Cdc42 previously (Itoh, R.E., K. Kurokawa, Y. Ohba, H. Yoshizaki, N. Mochizuki, and M. Matsuda. 2002. Mol. Cell. Biol. 22:6582–6591). Here, we added two types of probes for RhoA. One is to monitor the activity balance between guanine nucleotide exchange factors and GTPase-activating proteins, and another is to monitor the level of GTP-RhoA. Using these FRET probes, we imaged the activities of Rho-family GTPases during the cell division of HeLa cells. The activities of RhoA, Rac1, and Cdc42 were high at the plasma membrane in interphase, and decreased rapidly on entry into M phase. From after anaphase, the RhoA activity increased at the plasma membrane including cleavage furrow. Rac1 activity was suppressed at the spindle midzone and increased at the plasma membrane of polar sides after telophase. Cdc42 activity was suppressed at the plasma membrane and was high at the intracellular membrane compartments during cytokinesis. In conclusion, we could use the FRET-based probes to visualize the complex spatio-temporal regulation of Rho-family GTPases during cell division.
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spelling pubmed-21727912008-05-01 Activity of Rho-family GTPases during cell division as visualized with FRET-based probes Yoshizaki, Hisayoshi Ohba, Yusuke Kurokawa, Kazuo Itoh, Reina E. Nakamura, Takeshi Mochizuki, Naoki Nagashima, Kazuo Matsuda, Michiyuki J Cell Biol Article Rho-family GTPases regulate many cellular functions. To visualize the activity of Rho-family GTPases in living cells, we developed fluorescence resonance energy transfer (FRET)–based probes for Rac1 and Cdc42 previously (Itoh, R.E., K. Kurokawa, Y. Ohba, H. Yoshizaki, N. Mochizuki, and M. Matsuda. 2002. Mol. Cell. Biol. 22:6582–6591). Here, we added two types of probes for RhoA. One is to monitor the activity balance between guanine nucleotide exchange factors and GTPase-activating proteins, and another is to monitor the level of GTP-RhoA. Using these FRET probes, we imaged the activities of Rho-family GTPases during the cell division of HeLa cells. The activities of RhoA, Rac1, and Cdc42 were high at the plasma membrane in interphase, and decreased rapidly on entry into M phase. From after anaphase, the RhoA activity increased at the plasma membrane including cleavage furrow. Rac1 activity was suppressed at the spindle midzone and increased at the plasma membrane of polar sides after telophase. Cdc42 activity was suppressed at the plasma membrane and was high at the intracellular membrane compartments during cytokinesis. In conclusion, we could use the FRET-based probes to visualize the complex spatio-temporal regulation of Rho-family GTPases during cell division. The Rockefeller University Press 2003-07-21 /pmc/articles/PMC2172791/ /pubmed/12860967 http://dx.doi.org/10.1083/jcb.200212049 Text en Copyright © 2003, The Rockefeller University Press 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 Article
Yoshizaki, Hisayoshi
Ohba, Yusuke
Kurokawa, Kazuo
Itoh, Reina E.
Nakamura, Takeshi
Mochizuki, Naoki
Nagashima, Kazuo
Matsuda, Michiyuki
Activity of Rho-family GTPases during cell division as visualized with FRET-based probes
title Activity of Rho-family GTPases during cell division as visualized with FRET-based probes
title_full Activity of Rho-family GTPases during cell division as visualized with FRET-based probes
title_fullStr Activity of Rho-family GTPases during cell division as visualized with FRET-based probes
title_full_unstemmed Activity of Rho-family GTPases during cell division as visualized with FRET-based probes
title_short Activity of Rho-family GTPases during cell division as visualized with FRET-based probes
title_sort activity of rho-family gtpases during cell division as visualized with fret-based probes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2172791/
https://www.ncbi.nlm.nih.gov/pubmed/12860967
http://dx.doi.org/10.1083/jcb.200212049
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