Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 813K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JCB Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Komarova, Y. A. Articles by Borisy, G. G. Search for Related Content PubMed PubMed Citation Articles by Komarova, Y. A. Articles by Borisy, G. G. Related Collections Related Article Social Bookmarking                      What's this?

¹ Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, IL 60611
² Department of Cell Biology and Genetics, Erasmus University, 3015 GE Rotterdam, Netherlands

Address correspondence to Yulia Komarova, Dept. of Cell and Molecular Biology, Northwestern University Medical School, 303 E. Chicago Ave., Chicago, IL 60611-3008. Tel.: (312) 503-2854. Fax: (312) 503-7912. E-mail: y-komarova{at}northwestern.edu

The role of plus end–tracking proteins in regulating microtubule (MT) dynamics was investigated by expressing a dominant negative mutant that removed endogenous cytoplasmic linker proteins (CLIPs) from MT plus ends. In control CHO cells, MTs exhibited asymmetric behavior: MTs persistently grew toward the plasma membrane and displayed frequent fluctuations of length near the cell periphery. In the absence of CLIPs, the microtubule rescue frequency was reduced by sevenfold. MT behavior became symmetrical, consisting of persistent growth and persistent shortening. Removal of CLIPs also caused loss of p150^Glued but not CLIP-associating protein (CLASP2) or EB1. This result raised the possibility that the change in dynamics was a result of the loss of either CLIPs or p150^Glued. To distinguish between these possibilities, we performed rescue experiments. Normal MT dynamics were restored by expression of the CLIP-170 head domain, but p150^Glued was not recruited back to MT plus ends. Expression of p150^Glued head domain only partially restored MT dynamics. We conclude that the CLIP head domain is sufficient to alter MT dynamics either by itself serving as a rescue factor or indirectly by recruiting a rescue factor. By promoting a high rescue frequency, CLIPs provide a mechanism by which MT plus ends may be concentrated near the cell margin.

Key Words: plus end–tracking proteins; p150^Glued; cytoskeleton; dynamics; mammalian cell culture

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

Related Article

CLIPs to the rescue

Alan W. Dove
J. Cell Biol. 2002 159: 536. [Full Text] [PDF]

This article has been cited by other articles:

• Lewkowicz, E., Herit, F., Le Clainche, C., Bourdoncle, P., Perez, F., Niedergang, F. (2008). The microtubule-binding protein CLIP-170 coordinates mDia1 and actin reorganization during CR3-mediated phagocytosis. JCB 183: 1287-1298 [Abstract] [Full Text]  
• Odell, G. M., Foe, V. E. (2008). An agent-based model contrasts opposite effects of dynamic and stable microtubules on cleavage furrow positioning. JCB 183: 471-483 [Abstract] [Full Text]  
• Purro, S. A., Ciani, L., Hoyos-Flight, M., Stamatakou, E., Siomou, E., Salinas, P. C. (2008). Wnt Regulates Axon Behavior through Changes in Microtubule Growth Directionality: A New Role for Adenomatous Polyposis Coli. J. Neurosci. 28: 8644-8654 [Abstract] [Full Text]  
• Zovko, S., Abrahams, J. P., Koster, A. J., Galjart, N., Mommaas, A. M. (2008). Microtubule Plus-End Conformations and Dynamics in the Periphery of Interphase Mouse Fibroblasts. Mol. Biol. Cell 19: 3138-3146 [Abstract] [Full Text]  
• Helmstaedt, K., Laubinger, K., Vosskuhl, K., Bayram, O., Busch, S., Hoppert, M., Valerius, O., Seiler, S., Braus, G. H. (2008). The Nuclear Migration Protein NUDF/LIS1 Forms a Complex with NUDC and BNFA at Spindle Pole Bodies. Eukaryot Cell 7: 1041-1052 [Abstract] [Full Text]  
• Dragestein, K. A., van Cappellen, W. A., van Haren, J., Tsibidis, G. D., Akhmanova, A., Knoch, T. A., Grosveld, F., Galjart, N. (2008). Dynamic behavior of GFP-CLIP-170 reveals fast protein turnover on microtubule plus ends. JCB 180: 729-737 [Abstract] [Full Text]  
• Binker, M. G., Zhao, D. Y., Pang, S. J. Y., Harrison, R. E. (2007). Cytoplasmic Linker Protein-170 Enhances Spreading and Phagocytosis in Activated Macrophages by Stabilizing Microtubules. J. Immunol. 179: 3780-3791 [Abstract] [Full Text]  
• Eng, E. W., Bettio, A., Ibrahim, J., Harrison, R. E. (2007). MTOC Reorientation Occurs during Fc{gamma}R-mediated Phagocytosis in Macrophages. Mol. Biol. Cell 18: 2389-2399 [Abstract] [Full Text]  
• Gregoretti, I. V., Margolin, G., Alber, M. S., Goodson, H. V. (2006). Insights into cytoskeletal behavior from computational modeling of dynamic microtubules in a cell-like environment. J. Cell Sci. 119: 4781-4788 [Abstract] [Full Text]  
• Peris, L., Thery, M., Faure, J., Saoudi, Y., Lafanechere, L., Chilton, J. K., Gordon-Weeks, P., Galjart, N., Bornens, M., Wordeman, L., Wehland, J., Andrieux, A., Job, D. (2006). Tubulin tyrosination is a major factor affecting the recruitment of CAP-Gly proteins at microtubule plus ends. JCB 174: 839-849 [Abstract] [Full Text]  
• Vallee, R. B., Tsai, J.-W. (2006). The cellular roles of the lissencephaly gene LIS1, and what they tell us about brain development. Genes Dev. 20: 1384-1393 [Full Text]  
• Stehbens, S. J., Paterson, A. D., Crampton, M. S., Shewan, A. M., Ferguson, C., Akhmanova, A., Parton, R. G., Yap, A. S. (2006). Dynamic microtubules regulate the local concentration of E-cadherin at cell-cell contacts. J. Cell Sci. 119: 1801-1811 [Abstract] [Full Text]  
• Kita, K., Wittmann, T., Nathke, I. S., Waterman-Storer, C. M. (2006). Adenomatous Polyposis Coli on Microtubule Plus Ends in Cell Extensions Can Promote Microtubule Net Growth with or without EB1. Mol. Biol. Cell 17: 2331-2345 [Abstract] [Full Text]  
• Efimov, V. P., Zhang, J., Xiang, X. (2006). CLIP-170 Homologue and NUDE Play Overlapping Roles in NUDF Localization in Aspergillus nidulans. Mol. Biol. Cell 17: 2021-2034 [Abstract] [Full Text]  
• Xiang, X. (2006). A +TIP for a smooth trip. JCB 172: 651-654 [Abstract] [Full Text]  
• Folker, E. S., Baker, B. M., Goodson, H. V. (2005). Interactions between CLIP-170, Tubulin, and Microtubules: Implications for the Mechanism of CLIP-170 Plus-End Tracking Behavior. Mol. Biol. Cell 16: 5373-5384 [Abstract] [Full Text]  
• Komarova, Y., Lansbergen, G., Galjart, N., Grosveld, F., Borisy, G. G., Akhmanova, A. (2005). EB1 and EB3 Control CLIP Dissociation from the Ends of Growing Microtubules. Mol. Biol. Cell 16: 5334-5345 [Abstract] [Full Text]  
• Wu, X. S., Tsan, G. L., Hammer, J. A. III (2005). Melanophilin and myosin Va track the microtubule plus end on EB1. JCB 171: 201-207 [Abstract] [Full Text]  
• Akhmanova, A., Mausset-Bonnefont, A.-L., van Cappellen, W., Keijzer, N., Hoogenraad, C. C., Stepanova, T., Drabek, K., van der Wees, J., Mommaas, M., Onderwater, J., van der Meulen, H., Tanenbaum, M. E., Medema, R. H., Hoogerbrugge, J., Vreeburg, J., Uringa, E.-J., Grootegoed, J. A., Grosveld, F., Galjart, N. (2005). The microtubule plus-end-tracking protein CLIP-170 associates with the spermatid manchette and is essential for spermatogenesis. Genes Dev. 19: 2501-2515 [Abstract] [Full Text]  
• Dzhindzhev, N. S., Rogers, S. L., Vale, R. D., Ohkura, H. (2005). Distinct mechanisms govern the localisation of Drosophila CLIP-190 to unattached kinetochores and microtubule plus-ends. J. Cell Sci. 118: 3781-3790 [Abstract] [Full Text]  
• Wittmann, T., Waterman-Storer, C. M. (2005). Spatial regulation of CLASP affinity for microtubules by Rac1 and GSK3{beta} in migrating epithelial cells. JCB 169: 929-939 [Abstract] [Full Text]  
• Helmke, B. P. (2005). Molecular Control of Cytoskeletal Mechanics by Hemodynamic Forces. Physiology 20: 43-53 [Abstract] [Full Text]  
• Mimori-Kiyosue, Y., Grigoriev, I., Lansbergen, G., Sasaki, H., Matsui, C., Severin, F., Galjart, N., Grosveld, F., Vorobjev, I., Tsukita, S., Akhmanova, A. (2005). CLASP1 and CLASP2 bind to EB1 and regulate microtubule plus-end dynamics at the cell cortex. JCB 168: 141-153 [Abstract] [Full Text]  
• Kodama, A., Lechler, T., Fuchs, E. (2004). Coordinating cytoskeletal tracks to polarize cellular movements. JCB 167: 203-207 [Abstract] [Full Text]  
• Lansbergen, G., Komarova, Y., Modesti, M., Wyman, C., Hoogenraad, C. C., Goodson, H. V., Lemaitre, R. P., Drechsel, D. N., van Munster, E., Gadella, T. W.J. Jr., Grosveld, F., Galjart, N., Borisy, G. G., Akhmanova, A. (2004). Conformational changes in CLIP-170 regulate its binding to microtubules and dynactin localization. JCB 0: jcb.200402082-12 [Abstract] [Full Text]  
• Lallemand-Breitenbach, V., Quesnoit, M., Braun, V., El Marjou, A., Pous, C., Goud, B., Perez, F. (2004). CLIPR-59 Is a Lipid Raft-associated Protein Containing a Cytoskeleton-associated Protein Glycine-rich Domain (CAP-Gly) That Perturbs Microtubule Dynamics. J. Biol. Chem. 279: 41168-41178 [Abstract] [Full Text]  
• Liu, Q., Zuo, J., Pierce, E. A. (2004). The Retinitis Pigmentosa 1 Protein Is a Photoreceptor Microtubule-Associated Protein. J. Neurosci. 24: 6427-6436 [Abstract] [Full Text]  
• Sedbrook, J. C., Ehrhardt, D. W., Fisher, S. E., Scheible, W.-R., Somerville, C. R. (2004). The Arabidopsis SKU6/SPIRAL1 Gene Encodes a Plus End-Localized Microtubule-Interacting Protein Involved in Directional Cell Expansion. Plant Cell 16: 1506-1520 [Abstract] [Full Text]  
• Wittmann, T., Bokoch, G. M., Waterman-Storer, C. M. (2004). Regulation of Microtubule Destabilizing Activity of Op18/Stathmin Downstream of Rac1. J. Biol. Chem. 279: 6196-6203 [Abstract] [Full Text]  
• Lee, M., Lee, S., Zadeh, A. D., Kolodziej, P. A. (2003). Distinct sites in E-cadherin regulate different steps in Drosophila tracheal tube fusion. Development 130: 5989-5999 [Abstract] [Full Text]  
• Mimori-Kiyosue, Y., Tsukita, S. (2003). "Search-and-Capture" of Microtubules through Plus-End-Binding Proteins (+TIPs). J Biochem 134: 321-326 [Abstract] [Full Text]  
• Stepanova, T., Slemmer, J., Hoogenraad, C. C., Lansbergen, G., Dortland, B., De Zeeuw, C. I., Grosveld, F., van Cappellen, G., Akhmanova, A., Galjart, N. (2003). Visualization of Microtubule Growth in Cultured Neurons via the Use of EB3-GFP (End-Binding Protein 3-Green Fluorescent Protein). J. Neurosci. 23: 2655-2664 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents