Uropathogenic Escherichia coli (UPEC) is the most common cause of urinary tract infection, a disease of major significance to human health and increasingly associated with antibiotic resistance. UPEC possess multiple transcriptional regulators that facilitate its capacity to adapt to different environments and stresses. One such regulator belonging to the widespread MarR family, MprA, controls capsule and flagella expression, as well as expression of the EmrAB-TolC drug efflux pump. While MprA provides an intriguing link between the coordinated expression of virulence determinants and antibiotic resistance, its regulon and mechanisms of transcriptional control remain to be elucidated. Here, we employed transcriptome analysis (RNA-Seq) in conjunction with complementary protein-DNA mapping (ChIP-Seq) to define the MprA regulon in UPEC. Genes that were both directly regulated and differentially expressed were validated using promoter-reporter plasmids. Electrophoretic mobility shift assays, in combination with bioinformatic motif predictions, subsequently facilitated the identification of MprA binding sites in the promoter regions of select genes. Two novel targets were validated at the protein and phenotypic levels using western blotting and growth kinetics. Transcriptome analysis identified 66 differentially expressed target genes of MprA, including 35 previously undescribed genes. Of these 66 target genes, ChIP-Seq determined that 37 were directly regulated by MprA, and these results were validated using promoter-reporter analyses. The novel MprA-regulated targets included a gene encoding a common lipoprotein as well as genes involved in sialic acid catabolism, which were confirmed at the protein and phenotypic levels, respectively. Taken together, we have elucidated the regulon and protein-DNA interactions of MprA, a global UPEC regulator that coordinates the expression of both virulence and antibiotic resistance determinants in this clinically important antibiotic resistant pathogen.