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Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast
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Mps1 is SUMO-modified during the cell cycle.

Agnese Restuccia1, Feikun Yang2, Changyan Chen3

  • 1Division of Virus-Associated Carcinogenesis, German Cancer Research Center, Heidelberg, Germany.

Oncotarget
|December 18, 2015
PubMed
Summary
This summary is machine-generated.

Mps1 protein sumoylation is crucial for its function in mitosis. This modification affects Mps1

Keywords:
BubR1Mps1mitosissumoylation

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Mps1 (Monastrol-Plk1-binding sequence 1) is a kinase regulating the spindle assembly checkpoint and chromosome-microtubule attachment during mitosis.
  • The precise molecular mechanisms governing Mps1 protein levels and activity throughout the cell cycle are not fully understood.
  • Sumoylation (Small Ubiquitin-like Modifier conjugation) is a post-translational modification critical for mitotic progression.

Purpose of the Study:

  • To investigate whether Mps1 undergoes SUMO modification.
  • To determine the impact of sumoylation on Mps1 activity during mitosis.
  • To elucidate the role of sumoylation in Mps1's cellular localization and interactions.

Main Methods:

  • Western blotting using antibodies against SUMO-1 and SUMO-2 to detect Mps1 sumoylation.
  • Site-directed mutagenesis to identify key lysine residues involved in Mps1 sumoylation.
  • Immunofluorescence microscopy to assess Mps1 kinetochore localization.
  • Co-immunoprecipitation assays to analyze Mps1 interactions with BubR1.

Main Results:

  • Mps1 was found to be sumoylated by both SUMO-1 and SUMO-2 in asynchronous and mitotic cell populations.
  • Specific lysine residues (K71, K287, K367, K471) were identified as essential for Mps1 sumoylation.
  • Sumoylation was shown to influence Mps1's kinetochore localization, impacting mitotic progression.
  • SUMO-resistant Mps1 mutants exhibited enhanced interaction with BubR1 compared to wild-type Mps1.

Conclusions:

  • Mps1 undergoes significant SUMO modification during mitosis.
  • Sumoylation is essential for regulating Mps1's function, including its localization and interactions with key mitotic regulators like BubR1.
  • These findings provide critical insights into the post-translational regulation of Mps1 during cell division.