RecA protein plays a central role in homologous recombination and DNA repair. RecX, a gene directly downstream in Escherichia coli and some other bacterial species, down regulates it. However, the precise mechanism of regulation of RecA by RecX is not known. In order to study the function of RecX in the highly radioresistant bacterium Deinococcus radiodurans, null and overexpression strains were constructed. Our data demonstrates that RecX represses radiation resistance, DNA recombination and consequent genome instability in the stationary phase bacteria. Further biochemical analyses reveal that RecX not only down regulates recA transcription, but also directly inhibits RecA activities in vitro. These data suggests a dual negative regulatory control of RecX on RecA functions in D. radiodurans. © 2005 Elsevier B.V. All rights reserved.