Identification of cpxR as a positive regulator essential for expression of the Shigella sonnei virF gene.

virF is the master regulator which activates the virulence determinant genes of Shigella spp. such as ipaBCD and virG. We previously reported that expression of virF itself is regulated in a pH-dependent manner and that cpxA, a sensor of a two-component regulatory system, is involved in this regulation (S. Nakayama and H. Watanabe, J. Bacteriol. 177:5062-5069, 1995). Disruption of cpxR, which has been thought to be the cognate response regulator of cpxA (J. Dong, S. Iuchi, H.-S. Kwan, Z. Lue, and E. C. C. Lin, Gene 136:227-230, 1993), abolished virF expression almost completely. Purified CpxR bound directly to the upstream region of virF. Binding capacity was enhanced when CpxR was phosphorylated by coincubation with acetyl phosphate in vitro. Furthermore, we observed that phosphorylated CpxR could activate virF transcription in vitro. These results clearly indicated that CpxR was an essential activator for virF expression and strongly suggested that the binding of phosphorylated CpxR to the target site upstream of the virF gene induced a direct activation of virF transcription.

A simple technique for experimental hepatic vein catheterization in swine.

Hepatic vein catheterization is a valuable technique in studies of hepatic physiology and metabolism. A new technique for hepatic vein catheterization in swine is described which avoids fluoroscopy, incision, or puncture of the hepatic parenchyma. Experience with this new technique in over 40 studies of young pigs has confirmed the reliability of the technique. Management of hepatic vein catheters after insertion and potential sources of error in hepatic venous sampling are discussed.