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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...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
2003
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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. |
format | Text |
id | pubmed-2172791 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2003 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
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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