Conformational Changes in CLIP-170 Regulate Its Binding to Microtubules and Dynactin Localization

Cytoplasmic linker protein (CLIP)-170, CLIP-115, and the dynactin subunit p150Gluedare structurally related proteins, which associate specifically with the ends of growing microtubules (MTs). Here, we show that down-regulation of CLIP-170 by RNA interference results in a strongly reduced accumulatio...

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Bibliographic Details
Published in:The Journal of cell biology 2004-09, Vol.166 (7), p.1003-1014
Main Authors: Lansbergen, Gideon, Komarova, Yulia, Modesti, Mauro, Wyman, Claire, Hoogenraad, Casper C., Goodson, Holly V., Lemaitre, Régis P., Drechsel, David N., van Munster, Erik, Theodorus W. J. Gadella, Jr, Grosveld, Frank, Galjart, Niels, Borisy, Gary G., Akhmanova, Anna
Format: Article
Language:English
Subjects:
Amino Acid Motifs - genetics
Animals
Antibodies
Binding sites
Binding Sites - genetics
Cell biology
Cells
Cellular biology
Changes
CHO cells
COS Cells
Down-Regulation - genetics
Dynactin Complex
Fluorescence
HeLa cells
Microtubule-Associated Proteins - genetics
Microtubule-Associated Proteins - metabolism
Microtubules
Microtubules - genetics
Microtubules - metabolism
Microtubules - ultrastructure
Neoplasm Proteins
Protein Binding - genetics
Protein Conformation
Protein Structure, Tertiary - genetics
Protein Subunits - genetics
Protein Subunits - metabolism
Proteins
RNA Interference
Small interfering RNA
Zinc
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Summary:Cytoplasmic linker protein (CLIP)-170, CLIP-115, and the dynactin subunit p150Gluedare structurally related proteins, which associate specifically with the ends of growing microtubules (MTs). Here, we show that down-regulation of CLIP-170 by RNA interference results in a strongly reduced accumulation of dynactin at the MT tips. The NH2terminus of p150Gluedbinds directly to the COOH terminus of CLIP-170 through its second metal-binding motif. p150Gluedand LIS1, a dynein-associating protein, compete for the interaction with the CLIP-170 COOH terminus, suggesting that LIS1 can act to release dynactin from the MT tips. We also show that the NH2-terminal part of CLIP-170 itself associates with the CLIP-170 COOH terminus through its first metal-binding motif. By using scanning force microscopy and fluorescence resonance energy transfer-based experiments we provide evidence for an intramolecular interaction between the NH2and COOH termini of CLIP-170. This interaction interferes with the binding of the CLIP-170 to MTs. We propose that conformational changes in CLIP-170 are important for binding to dynactin, LIS1, and the MT tips.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.200402082