Iron-sensitive RNA regulation by poly C binding proteins [RIP-Seq]

Iron is important for normal cellular function and posttranscriptional mechanisms regulate the cellular response to iron deficiency. In this study we explore how RNA binding proteins, PCBP1 and PCBP2, exhibit iron-sensitive RNA regulation. Overall design: K562 cells were induced (with doxycycline) to expressed either a Flag-tagged PCBP1, wildtype (WT), a non-iron binding mutant (?Fe), a non-RNA binding mutant (?RNA), or Flag-tagged-GFP. Cells were then treated overnight with DMSO (control) or 10 µM iron chelator (21H7). With two biological replicates, cells were subjected to formaldehyde crosslinking and immunoprecipitation using anti-Flag antibodies. Bound RNA was isolated and sequenced along side inputs.