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An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C
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m6Am methyltransferase PCIF1 negatively regulates ciliation by inhibiting BICD2 expression

Shanshan Xie1, Wenjun Kuang2, Mengzhe Guo3

  • 1Children's Hospital, National Clinical Research Center for Child Health, Zhejiang University School of Medicine, Hangzhou, China.

The Journal of Cell Biology
|March 25, 2024
PubMed

Insights

Phosphorylated CTD interacting factor 1 (PCIF1) regulates ciliogenesis by adding m6Am RNA modifications. PCIF1

Area of Science:

  • Molecular Biology
  • Cell Biology
  • RNA Modifications

Background:

  • N6, 2'-O-dimethyladenosine (m6Am) is a prevalent epitranscriptomic mark.
  • The biological roles of m6Am and its catalyzing enzyme, PCIF1, are not fully understood.

Purpose of the Study:

  • To investigate the function of PCIF1-dependent m6Am modification in RPE-1 cells.
  • To elucidate the role of PCIF1 in regulating ciliogenesis.

Main Methods:

  • Quantitative proteomic analysis
  • RNA sequencing
  • LC-MS analysis
  • Cell culture and manipulation (siRNA, knockdown)

Main Results:

  • PCIF1 acts as a negative regulator of ciliogenesis via its m6Am methyltransferase activity.
  • BICD2 is identified as a downstream target of PCIF1, with its levels inversely correlated with PCIF1 activity.
  • PCIF1-mediated m6Am modification of BICD2 mRNA reduces its stability and translation efficiency, impacting ciliation.

Conclusions:

  • PCIF1-dependent m6Am modification is crucial for regulating ciliogenesis.
  • The PCIF1-BICD2 axis represents a novel regulatory pathway in ciliation.

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