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The two-component regulatory system, composed ofvirA andvirG , is indispensable for transcription of virulence genes withinAgrobacterium tumefaciens . However,virA andvirG are insufficient to activate transcription from virulence gene promoters withinEscherichia coli cells, indicating a requirement for additionalA. tumefaciens genes. In a search for these additional genes, we have identified therpoA gene, encoding the α subunit of RNA polymerase (RNAP), which confers significant expression of avirB promoter (virBp )::lacZ fusion inE. coli in the presence of an active transcriptional regulatorvirG gene. We conducted in vitro transcription assays using either reconstitutedE. coli RNAP or hybrid RNAP in which the α subunit was derived fromA. tumefaciens . The two forms of RNAP were equally efficient in transcription from a ς70 -dependentE. coli gal P1 promoter; however, only the hybrid RNAP was able to transcribevirBp in avirG -dependent manner. In addition, we provide evidence that the α subunit fromA. tumefaciens , but not fromE. coli , is able to interact with the VirG protein. These data suggest that transcription of virulence genes requires specific interaction between VirG and the α subunit ofA. tumefaciens and that the α subunit fromE. coli is unable to effectively interact with the VirG protein. This work provides the basis for future studies designed to examinevir gene expression as well as the T-DNA transfer process inE. coli.

journal of bacteriology

Transcriptional Activation of Agrobacterium tumefaciens Virulence Gene Promoters in Escherichia coli Requires the A. tumefaciens rpoA Gene, Encoding the Alpha Subunit of RNA Polymerase