Mimicking Ndc80 phosphorylation triggers spindle assembly checkpoint signalling
| Autor: | Thomas Ruppert, Stefan Kemmler, Jennifer Ortiz, Jens Pfannstiel, Johannes Lechner, Maria Knapp, Manuel Stach |
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| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
Saccharomyces cerevisiae Proteins
Recombinant Fusion Proteins Molecular Sequence Data Saccharomyces cerevisiae Spindle Apparatus Biology Protein Serine-Threonine Kinases Microtubules General Biochemistry Genetics and Molecular Biology Article chemistry.chemical_compound Aurora Kinases Animals Amino Acid Sequence Nuclear protein Phosphorylation Protein kinase A Kinetochores Molecular Biology Aspartic Acid Alanine General Immunology and Microbiology Kinetochore General Neuroscience Cell Cycle Intracellular Signaling Peptides and Proteins Nuclear Proteins Protein-Tyrosine Kinases Cell biology NDC80 Genes cdc Spindle checkpoint Nocodazole chemistry Signal transduction Protein Kinases Signal Transduction |
| Popis: | The protein kinase Mps1 is, among others, essential for the spindle assembly checkpoint (SAC). We found that Saccharomyces cerevisiae Mps1 interacts physically with the N-terminal domain of Ndc80 (Ndc80(1-257)), a constituent of the Ndc80 kinetochore complex. Furthermore, Mps1 effectively phosphorylates Ndc80(1-257) in vitro and facilitates Ndc80 phosphorylation in vivo. Mutating 14 of the phosphorylation sites to alanine results in compromised checkpoint signalling upon nocodazole treatment of mutants. Mutating the identical sites to aspartate (to simulate constitutive phosphorylation) causes a metaphase arrest with wild-type-like bipolar kinetochore-microtubule attachment. This arrest is due to a constitutively active SAC and consequently the inviable aspartate mutant can be rescued by disrupting SAC signalling. Therefore, we conclude that a putative Mps1-dependent phosphorylation of Ndc80 is important for SAC activation at kinetochores. |
| Databáze: | OpenAIRE |
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