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Iron-sensitive RNA regulation by poly C-binding proteins.

Grant A Goda1,2, Kwame Forbes2,3, Michael E Sullivan2

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Poly C-binding proteins (PCBPs) are crucial for managing cellular iron. This study reveals how low iron levels impact PCBP-regulated gene expression and alternative splicing, identifying PCBPs as key players in iron-sensitive RNA regulation.

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Area of Science:

  • Molecular Biology
  • Cellular Biology
  • RNA Biology

Background:

  • Iron is vital for cellular function, with homeostasis maintained by posttranscriptional mechanisms.
  • Poly C-binding proteins (PCBPs) are essential RNA-binding proteins involved in RNA processing and iron transport.
  • The influence of cellular iron on the PCBP-regulated transcriptome remains largely uncharacterized.

Purpose of the Study:

  • To globally evaluate the impact of cellular iron levels on the PCBP-directed transcriptome.
  • To identify genes with iron-sensitive splicing regulation mediated by PCBPs.
  • To elucidate the role of PCBPs in iron-mediated posttranscriptional regulation.

Main Methods:

  • Transcriptome-wide analysis of gene expression and alternative splicing (AS) under low iron conditions.
  • Comparison of AS patterns directed by PCBP1 and PCBP2 with the iron-sensitive transcriptome.
  • Gene knockdown experiments to assess the role of PCBPs in iron-induced splicing changes.
  • RNA immunoprecipitation assays to study PCBP1-RNA interactions.

Main Results:

  • Low iron availability induces broad transcriptome changes, including significant alterations in alternative splicing.
  • Specific genes exhibit iron-sensitive splicing regulation mediated by PCBP1 and PCBP2.
  • Knockdown of PCBPs attenuates splicing changes induced by iron chelation.
  • Iron chelation or mutation of PCBP1's iron-binding sites increases its association with RNA.

Conclusions:

  • Cellular iron levels broadly influence RNA regulation, particularly alternative splicing.
  • PCBP1 and PCBP2 are identified as critical splicing factors in the cellular response to iron.
  • This study highlights a novel mechanism of iron-sensitive RNA regulation involving PCBPs.