Hypoxia‐Inducible Factor‐1 (HIF‐1) has been largely studied for its role in cell survival in hypoxic conditions. The regulation of HIF‐1 is a complex process and involves a number of molecules and pathways. Among these mechanisms a direct regulatory role of reactive oxygen species (ROS) on HIF‐1 alpha subunit has received a great deal of attention and the existing body of literature includes many contradictory findings. Other intermediates such as nitric oxide (NO), specific microRNAs (miR), and transcriptional and post‐translational modification have also been implicated as players in ROS mediated HIF‐1a regulation. The focus of this review is to present the past conflicting evidence along with more recent findings in order to relate various aspects of this complex process. Aside from the direct role of ROS on HIF‐1a regulation under hypoxia and normoxia, we analyzed the effect of different sources and concentrations of NO and the interplay between superoxide (SO) and NO in this process. We also present findings on transcriptional and translational regulation of HIF‐1a via ROS and the interplay with microRNAs in this process. This review further provides insight on ERK and PI3K/AKT signaling as a common mechanism relating several pathways of ROS mediated HIF‐1a regulation. Ultimately further research and discovery regarding HIF‐1 regulation by oxidative stress is warranted for better understanding of disease development and potential therapeutics for pathologies such as cancer, inflammatory diseases, and ischemia‐reperfusion injury. J. Cell. Biochem. 116: 696–703, 2015. © 2014 Wiley Periodicals, Inc.