Mg2+ is the most abundant divalent cation in biological systems. It is required for ATP-mediated enzymatic reactions and as a stabilizer of ribosomes and membranes. The enteric bacterium Salmonella enterica serovar Typhimurium harbors three Mg2+ transporters and a regulatory system-termed PhoP/PhoQ-whose activity is regulated by the extracytoplasmic levels of Mg2+. We have determined that expression of the PhoP-activated Mg2+ transporter MgtA is also controlled by its 5'-untranslated region (5'UTR). The 5'UTR of the mgtA gene can adopt different stem-loop structures depending on the Mg2+ levels, which determine whether transcription reads through into the mgtA-coding region or stops within the 5'UTR. This makes the mgtA 5'UTR the first example of a cation-responding riboswitch. The initiation of mgtA transcription responds to extracytoplasmic Mg2+, and its elongation into the coding region to cytoplasmic Mg2+, which provides a singular example where the same ligand is sensed in different cellular compartments to regulate disparate steps in gene transcription. The PhoP-activated Mg2+ transporter mgtB is also regulated by Mg2+ in a strain lacking the Mg2+ sensor PhoQ, suggesting the presence of additional Mg2+-responding devices.