Many intracellular bacterial pathogens use type IV secretion systems to deliver effector molecules and subvert the eukaryotic host cell defenses. The genus Brucella comprises facultative intracellular bacteria that cause brucellosis, a disease affecting a wide range of mammals including humans. The virB operon codes for a type IV secretion system that plays a central role in intracellular survival and replication of Brucella within the host. Expression of the virB genes is under the control of various transcription factors that allow this system to respond to different types of environmental signals, and display binding site structures and arrangements that define the intrinsic complexity of the virB promoter. This review focuses on summarizing the current state of research concerning regulation of the Brucella virB operon, with special emphasis on describing the nature and function of the implicated regulatory elements and examining the involved protein-DNA interactions.