In bacteria-host interactions, competition for iron is critical for the outcome of the infection. As a result of its redox properties, this metal is essential for the growth and proliferation of most living organisms, including pathogenic bacteria. This metal is also potentially toxic, making the precise maintenance of iron homeostasis necessary for survival. Iron acquisition and storage control is mediated in most bacteria by the global ferric uptake regulator (Fur) and iron-responsive small regulatory non-coding RNAs (RyhB in the model organism Escherichia coli). While the role of these regulators in iron homeostasis is well documented in both pathogenic and non-pathogenic bacteria, many recent studies also demonstrate that these regulators are involved in the virulence of pathogenic bacteria. By sensing iron availability in the environment, Fur and RyhB are able to regulate, either directly or indirectly via other transcriptional regulators or modulation of intracellular iron concentration, many virulence determinants of pathogenic bacteria. Iron is thus both a nutritional and regulatory element, allowing bacteria to adapt to various host environments by adjusting expression of virulence factors. In this review, we present evidences that Fur and RyhB are the major regulators of this adaptation, as they are involved in diverse functions ranging from iron homeostasis to regulation of virulence by mediating key pathogen responses such as invasion of eukaryotic cells, toxin production, motility, quorum sensing, stress resistance or biofilm formation. Therefore, Fur and RyhB play a major role in regulating an adaptative response during bacterial infections, making them important targets in the fight against pathogenic bacteria.