We have identified two closely linked genes, pilA and pilB, which act in trans on the pilin promoter. pilA-pilB map downstream of expression loci pilE1 and opaE1 in the gonococcal chromosome. Subcloning data indicate that pilB acts negatively on the pilin promoter, and insertional inactivation of pilB results in hyperpiliated gonococci. A pilA clone activates the pilin promoter in Escherichia coli, and a pilA-/pilA+ heterodiploid gonococcus exhibits a P- phenotype. Our inability to obtain simple pilA- mutants strongly suggests that pilA is an essential gene in the gonococcus. In an in vitro coupled transcription/translation system, inserts spanning the pilA and pilB region direct the synthesis of two proteins of 40 and 58 kd. DNA sequence analysis shows that the pilA and pilB loci encode proteins of 38.6 kd (with a putative DNA binding domain) and 57.9 kd respectively. The pilA and pilB genes are in opposite orientation relative to each other, and the 5' ends of the two genes overlap. We discuss how these two loci may interact to control pilin expression in the gonococcus.