Phosphorylation controls autoinhibition of cytoplasmic linker protein-170
Molecular Biology of the Cell (Print) , Volume 21 - Issue 15 p. 2661- 2673
Cytoplasmic linker protein (CLIP)-170 is a microtubule (MT) plus-end-tracking protein that regulates MT dynamics and links MT plus ends to different intracellular structures. We have shown previously that intramolecular association between the N and C termini results in autoinhibition of CLIP-170, thus altering its binding to MTs and the dynactin subunit p150Glued(J. Cell Biol. 2004: 166, 1003-1014). In this study, we demonstrate that conformational changes in CLIP-170 are regulated by phosphorylation that enhances the affinity between the N- and C-terminal domains. By using site-directed mutagenesis and phosphoproteomic analysis, we mapped the phosphorylation sites in the third serine-rich region of CLIP-170. A phosphorylation-deficient mutant of CLIP-170 displays an "open" conformation and a higher binding affinity for growing MT ends and p150Gluedas compared with nonmutated protein, whereas a phosphomimetic mutant confined to the "folded back" conformation shows decreased MT association and does not interact with p150Glued. We conclude that phosphorylation regulates CLIP-170 conformational changes resulting in its autoinhibition.
|Molecular Biology of the Cell (Print)|
|Organisation||Erasmus MC: University Medical Center Rotterdam|
Lee, H.S, Komarova, Y, Nadezhdina, E, Anjum, R, Peloquin, J.G, Schober, J.M, … Borisy, G.G. (2010). Phosphorylation controls autoinhibition of cytoplasmic linker protein-170. Molecular Biology of the Cell (Print), 21(15), 2661–2673. doi:10.1091/mbc.E09-12-1036