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Combining Mitotic Cell Synchronization and High Resolution Confocal Microscopy to Study the Role of Multifunctional Cell Cycle Proteins During Mitosis
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YTHDF2 promotes mitotic entry and is regulated by cell cycle mediators.

Qili Fei1,2,3, Zhongyu Zou1,2, Ian A Roundtree1,2,4,5,6

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The N6-methyladenosine (m6A) reader YTHDF2 promotes mRNA decay for cell cycle progression. Its stability, regulated by CDK1, forms a loop with WEE1 to ensure timely mitotic entry.

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • Cell Biology

Background:

  • N6-methyladenosine (m6A) is a crucial epitranscriptomic modification regulating mRNA fate.
  • The m6A reader protein YTHDF2 plays a role in mRNA decay, but its cell cycle function is not fully understood.

Purpose of the Study:

  • To investigate the role of YTHDF2 in cell cycle regulation.
  • To elucidate the mechanisms by which m6A modification influences cell cycle progression.

Main Methods:

  • Depletion of YTHDF2 in HeLa cells.
  • Analysis of cell cycle regulators, including WEE1.
  • Investigation of YTHDF2 protein stability and its regulation by CDK1.
  • Identification of E3 ubiquitin ligase components involved in YTHDF2 degradation.

Main Results:

  • YTHDF2 depletion delays mitotic entry by causing the accumulation of cell cycle inhibitors like WEE1.
  • WEE1 transcripts are m6A-modified and undergo YTHDF2-mediated decay.
  • YTHDF2 protein stability is regulated by CDK1 activity, forming a positive feedback loop with WEE1.
  • CUL1, CUL4A, DDB1, and SKP2 are identified as components of E3 ubiquitin ligase complexes targeting YTHDF2 for degradation.

Conclusions:

  • CDK1, YTHDF2, and WEE1 form a feedforward loop that promotes mitotic entry.
  • Cell cycle mediators dynamically regulate transcriptomic m6A modification, impacting cell cycle progression.
  • This study reveals a novel regulatory mechanism linking epitranscriptomics to cell cycle control.