In the oligotrophic freshwater bacterium Caulobacter crescentus, D-xylose induces expression of over 50 genes,including the xyl operon, which encodes key enzymes for xylose metabolism. The promoter (PxylX) controllingexpression of the xyl operon is widely used as a tool for inducible heterologous gene expression in C. crescentus. We show here that PxylX and at least one other promoter in the xylose regulon (PxylE) are controlled by theCC3065 (xylR) gene product, a LacI-type repressor. Electrophoretic gel mobility shift assays showed that operator binding by XylR is greatly reduced in the presence of D-xylose. The data support the hypothesis that there is a simple regulatory mechanism in which XylR obstructs xylose-inducible promoters in the absence of the sugar; the repressor is induced to release DNA upon binding D-xylose, thereby freeing the promoter for productive interaction with RNA polymerase. XylR also has an effect on glucose metabolism, as xylR mutantsexhibit reduced expression of the Entner-Doudoroff operon and their ability to utilize glucose as a sole carbon and energy source is compromised.