Sequence Determinants of a Microtubule Tip Localization Signal (MtLS)
| Autor: | José M. de Pereda, Indrani Sen, David Gfeller, Oliver Kortt, Rubén M. Buey, Dario Neri, Rudolf Volkmer, Renu Mohan, Olivier Michielin, Jörg Scheuermann, Ines Kretzschmar, Vincent Zoete, Anna Akhmanova, Dmitry B. Veprintsev, Michel O. Steinmetz |
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| Přispěvatelé: | Swiss National Science Foundation, European Commission, Federation of European Biochemical Societies, EMBO, Ministerio de Ciencia e Innovación (España) |
| Rok vydání: | 2012 |
| Předmět: |
Amino Acid Motifs
Protein domain Arabidopsis Protein Array Analysis Plasma protein binding Protein Sorting Signals Biology Microtubules Biochemistry 03 medical and health sciences 0302 clinical medicine Microtubule Schizosaccharomyces Humans Short linear motif Structural motif Cytoskeleton Molecular Biology 030304 developmental biology 0303 health sciences Arabidopsis Proteins Cell Biology 3. Good health Cell biology Microtubule Proteins Schizosaccharomyces pombe Proteins Molecular Biophysics 030217 neurology & neurosurgery Protein Binding |
| Zdroj: | The Journal of biological chemistry J. Biol. Chem. Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m112.373928 |
| Popis: | et al. Microtubule plus-end-tracking proteins (+TIPs) specifically localize to the growing plus-ends of microtubules to regulate microtubule dynamics and functions. A large group of +TIPs contain a short linear motif, SXIP, which is essential for them to bind to end-binding proteins (EBs) and target microtubule ends. The SXIP sequence site thus acts as a widespread microtubule tip localization signal (MtLS). Here we have analyzed the sequence-function relationship of a canonical MtLS. Using synthetic peptide arrays on membrane supports, we identified the residue preferences at each amino acid position of the SXIP motif and its surrounding sequence with respect to EB binding. We further developed an assay based on fluorescence polarization to assess the mechanism of the EB-SXIP interaction and to correlate EB binding and microtubule tip tracking of MtLS sequences from different +TIPs. Finally, we investigated the role of phosphorylation in regulating the EB-SXIP interaction. Together, our results define the sequence determinants of a canonical MtLS and provide the experimental data for bioinformatics approaches to carry out genome-wide predictions of novel +TIPs in multiple organisms. This work was supported by a Federation of European Biochemical Societies (FEBS) postdoctoral fellowship and a contract from the Juan de la Cierva program and a Marie Curie Career Integration grant (EB-SxIP; 293831) (to R. M. B.), by a Marie Curie IIF grant (MT-TIP Inhibitors; 253818) (to R. M.), by an EMBO grant (to D. G.), and by Swiss National Science Foundation Grants 31003A_122545 and 310030B_138659 (to M. O. S.) |
| Databáze: | OpenAIRE |
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