Iron (Fe) is an essential micronutrient for plant growth development and plays a key role in regulating numerous cellular processes. In rice, OsHRZ1, an Fe-binding ubiquitin ligase, is a putative sensor of Fe homeostasis that negatively regulates iron acquisition. Despite its apparent importance, only a single basic-Helix-Loop-Helix (bHLH) transcription factor, OsPRI1, has been identified as a direct target of OsHRZ1. In this study, we identified and functionally characterized OsPRI2 and OsPRI3, two paralogs of OsPRI1, observing that they directly interact with OsHRZ1. Additional analyses suggested that OsHRZ1 promotes the degradation of OsPRI2 and OsPRI3. The translocation of Fe from roots to shoots was impaired in plants with loss-of-function mutations in OsPRI2 or OsPRI3, causing the downregulation of Fe-deficiency-responsive genes. In contrast, overexpression of OsPRI2 and OsPRI3 promotes Fe accumulation and activates the expression of Fe-deficiency-responsive genes. We also provide evidence that OsPRI2 and OsPRI3 bind to the promoters of OsIRO2 and OsIRO3, two key regulators of Fe homeostasis. Moreover, OsPRI2 and OsPRI3 directly induce expression of the metal-nicotianamine transporter, OsYSL2, by associating with the promoter in response to Fe deficiency. Our results provide insights into the complex network regulating Fe homeostasis in rice.