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Dynamic behavior of GFP–CLIP-170 reveals fast protein turnover on microtubule plus ends
Microtubule (MT) plus end–tracking proteins (+TIPs) specifically recognize the ends of growing MTs. +TIPs are involved in diverse cellular processes such as cell division, cell migration, and cell polarity. Although +TIP tracking is important for these processes, the mechanisms underlying plus end s...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2265578/ https://www.ncbi.nlm.nih.gov/pubmed/18283108 http://dx.doi.org/10.1083/jcb.200707203 |
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author | Dragestein, Katharina A. van Cappellen, Wiggert A. van Haren, Jeffrey Tsibidis, George D. Akhmanova, Anna Knoch, Tobias A. Grosveld, Frank Galjart, Niels |
author_facet | Dragestein, Katharina A. van Cappellen, Wiggert A. van Haren, Jeffrey Tsibidis, George D. Akhmanova, Anna Knoch, Tobias A. Grosveld, Frank Galjart, Niels |
author_sort | Dragestein, Katharina A. |
collection | PubMed |
description | Microtubule (MT) plus end–tracking proteins (+TIPs) specifically recognize the ends of growing MTs. +TIPs are involved in diverse cellular processes such as cell division, cell migration, and cell polarity. Although +TIP tracking is important for these processes, the mechanisms underlying plus end specificity of mammalian +TIPs are not completely understood. Cytoplasmic linker protein 170 (CLIP-170), the prototype +TIP, was proposed to bind to MT ends with high affinity, possibly by copolymerization with tubulin, and to dissociate seconds later. However, using fluorescence-based approaches, we show that two +TIPs, CLIP-170 and end-binding protein 3 (EB3), turn over rapidly on MT ends. Diffusion of CLIP-170 and EB3 appears to be rate limiting for their binding to MT plus ends. We also report that the ends of growing MTs contain a surplus of sites to which CLIP-170 binds with relatively low affinity. We propose that the observed loss of fluorescent +TIPs at plus ends does not reflect the behavior of single molecules but is a result of overall structural changes of the MT end. |
format | Text |
id | pubmed-2265578 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-22655782008-08-25 Dynamic behavior of GFP–CLIP-170 reveals fast protein turnover on microtubule plus ends Dragestein, Katharina A. van Cappellen, Wiggert A. van Haren, Jeffrey Tsibidis, George D. Akhmanova, Anna Knoch, Tobias A. Grosveld, Frank Galjart, Niels J Cell Biol Research Articles Microtubule (MT) plus end–tracking proteins (+TIPs) specifically recognize the ends of growing MTs. +TIPs are involved in diverse cellular processes such as cell division, cell migration, and cell polarity. Although +TIP tracking is important for these processes, the mechanisms underlying plus end specificity of mammalian +TIPs are not completely understood. Cytoplasmic linker protein 170 (CLIP-170), the prototype +TIP, was proposed to bind to MT ends with high affinity, possibly by copolymerization with tubulin, and to dissociate seconds later. However, using fluorescence-based approaches, we show that two +TIPs, CLIP-170 and end-binding protein 3 (EB3), turn over rapidly on MT ends. Diffusion of CLIP-170 and EB3 appears to be rate limiting for their binding to MT plus ends. We also report that the ends of growing MTs contain a surplus of sites to which CLIP-170 binds with relatively low affinity. We propose that the observed loss of fluorescent +TIPs at plus ends does not reflect the behavior of single molecules but is a result of overall structural changes of the MT end. The Rockefeller University Press 2008-02-25 /pmc/articles/PMC2265578/ /pubmed/18283108 http://dx.doi.org/10.1083/jcb.200707203 Text en Copyright © 2008, 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 | Research Articles Dragestein, Katharina A. van Cappellen, Wiggert A. van Haren, Jeffrey Tsibidis, George D. Akhmanova, Anna Knoch, Tobias A. Grosveld, Frank Galjart, Niels Dynamic behavior of GFP–CLIP-170 reveals fast protein turnover on microtubule plus ends |
title | Dynamic behavior of GFP–CLIP-170 reveals fast protein turnover on microtubule plus ends |
title_full | Dynamic behavior of GFP–CLIP-170 reveals fast protein turnover on microtubule plus ends |
title_fullStr | Dynamic behavior of GFP–CLIP-170 reveals fast protein turnover on microtubule plus ends |
title_full_unstemmed | Dynamic behavior of GFP–CLIP-170 reveals fast protein turnover on microtubule plus ends |
title_short | Dynamic behavior of GFP–CLIP-170 reveals fast protein turnover on microtubule plus ends |
title_sort | dynamic behavior of gfp–clip-170 reveals fast protein turnover on microtubule plus ends |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2265578/ https://www.ncbi.nlm.nih.gov/pubmed/18283108 http://dx.doi.org/10.1083/jcb.200707203 |
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