Requirements for Protein Phosphorylation and the Kinase Activity of Polo-like Kinase 1 (Plk1) for the Kinetochore Function of Mitotic Arrest Deficiency Protein 1 (Mad1)
| Autor: | Yan Li, Michael D. Ward, Kerstin Haller, Kuan-Teh Jeang, Ya-Hui Chi, O. John Semmes |
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| Rok vydání: | 2008 |
| Předmět: |
Mad1
Mitosis Cell Cycle Proteins macromolecular substances Protein Serine-Threonine Kinases Biochemistry PLK1 MAP2K7 Molecular Basis of Cell and Developmental Biology Proto-Oncogene Proteins Humans Mass Screening Protein phosphorylation Phosphorylation RNA Small Interfering Kinase activity Kinetochores Protein kinase A Molecular Biology biology Cyclin-dependent kinase 2 Nuclear Proteins Cell Biology Autophagy-related protein 13 Cell biology biology.protein Chromatography Liquid HeLa Cells |
| Zdroj: | Journal of Biological Chemistry. 283:35834-35844 |
| ISSN: | 0021-9258 |
| Popis: | Mitotic arrest deficiency protein 1 (Mad1) is associated with microtubule-unattached kinetochores in mitotic cells and is a component of the spindle assembly checkpoint (SAC). Here, we have studied the phosphorylation of Mad1 and mapped using liquid chromatography-tandem mass spectrometry several phosphorylated amino acids in this protein. One phosphorylated residue, Thr680, was characterized to be important for the kinetochore localization of Mad1 and its SAC function. We also found that in mitotic cells Mad1 co-immunoprecipitated with Plk1. Depletion of cellular Plk1 using small interfering RNAs and inhibition of the kinase activity of Plk1 using a kinase-dead mutant or a small molecule inhibitor attenuated Mad1 phosphorylation and its association with kinetochores. Collectively, these findings indicate mechanistic roles contributed by protein phosphorylation and Plk1 to the SAC activity of Mad1. |
| Databáze: | OpenAIRE |
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