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Specific inhibition of NF-Y subunits triggers different cell proliferation defects

Paolo Benatti1, Diletta Dolfini, Alessandra Viganò

  • 1Dipartimento di Biologia, Università di Modena e Reggio Emilia, via Campi 213/D, 41125 Modena, Italy.

Nucleic Acids Research
|March 19, 2011
PubMed

Insights

The transcription factor NF-Y regulates cell cycle genes. Silencing NF-YA delays S-phase and causes DNA damage, while NF-YB loss prevents G2/M exit, revealing distinct subunit roles in cell cycle control.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Regulated gene expression is crucial for cell cycle progression.
  • The transcription factor NF-Y plays a key role in regulating cell cycle genes, especially those involved in the G2/M phase.

Purpose of the Study:

  • To investigate the distinct and common functions of NF-Y subunits (NF-YA, NF-YB, NF-YC) in cell cycle regulation.
  • To understand how the loss of individual NF-Y subunits affects cellular responses and gene expression.

Main Methods:

  • Silencing of NF-YA, NF-YB, and NF-YC using shRNAs in HCT116 cells.
  • Analysis of cell cycle progression, DNA damage, apoptosis, and gene expression.
  • Chromatin immunoprecipitation (ChIP) assays to assess NF-Y binding to target genes.

Main Results:

  • NF-YA depletion caused S-phase delay, DNA damage, and apoptosis, activating the replication checkpoint.
  • NF-YB depletion hindered G2/M exit without affecting S-phase progression.
  • Gene expression analysis revealed both common and distinct target genes for NF-YA and NF-YB.
  • NF-YA depletion was more effective than NF-YB in disrupting NF-Y binding to CCAAT promoters.

Conclusions:

  • Loss of individual NF-Y subunits leads to distinct cellular responses and cell cycle arrest pathways.
  • NF-Y subunits play critical but differential roles in maintaining cell cycle integrity.
  • Cellular perception of NF-Y subunit loss activates diverse signaling pathways and cell cycle blocks.

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