Abstract
A chimera with the green fluorescent protein (GFP) has been constructed to visualize the dynamic properties of the endosome-microtubule linker protein CLIP170 (GFP-CLIP170). GFP-CLIP170 binds in stretches along a subset of microtubule ends. These fluorescent stretches appear to move with the growing tips of microtubules at 0.15-0.4 microm/s, comparable to microtubule elongation in vivo. Analysis of speckles along dynamic GFP-CLIP170 stretches suggests that CLIP170 treadmills on growing microtubule ends, rather than being continuously transported toward these ends. Drugs affecting microtubule dynamics rapidly inhibit movement of GFP-CLIP170 dashes. We propose that GFP-CLIP170 highlights growing microtubule ends by specifically recognizing the structure of a segment of newly polymerized tubulin.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antineoplastic Agents / pharmacology
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Antineoplastic Agents, Phytogenic / pharmacology
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Chlorocebus aethiops
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Gene Expression / physiology
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Green Fluorescent Proteins
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HeLa Cells
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Humans
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Indicators and Reagents / metabolism
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Kinetics
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Microscopy, Fluorescence
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Microtubule-Associated Proteins / genetics*
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Microtubule-Associated Proteins / metabolism*
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Microtubules / drug effects
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Microtubules / metabolism*
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Neoplasm Proteins
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Nocodazole / pharmacology
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Paclitaxel / pharmacology
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Recombinant Fusion Proteins / metabolism
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Vero Cells
Substances
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Antineoplastic Agents
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Antineoplastic Agents, Phytogenic
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Indicators and Reagents
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Luminescent Proteins
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Microtubule-Associated Proteins
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Neoplasm Proteins
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Recombinant Fusion Proteins
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Green Fluorescent Proteins
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cytoplasmic linker protein 170
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Paclitaxel
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Nocodazole