KdpD and KdpE, proteins that control expression of the kdpABC operon, are members of the two-component sensor-effector class of regulators

Abstract

The Kdp system of Escherichia coli, a transport ATPase with high affinity for potassium, is expressed when turgor pressure is low. Expression requires KdpD, a 99-kDa membrane protein, and KdpE, a 25-kDa soluble cytoplasmic protein. (16,41,49). We suggest that phosphoKdpE is a positive effector of Kdp expression and that low turgor pressure causes KdpD to phosphorylate KdpE. MATERUILS AND METHODSDNA sequencing. The 1.7-kb EcoRI fragment of kdpD, cut from plasmid pWE1103 (35), and the 3-kb EcoRI-HindIII fragment carrying kdpE and a part of kdpD, cut from plasmid pDE14 (35), were cloned in both directions in M13 (M13uml8 and M13uml9; International Biotechnologies, Inc.). The resulting phages, M13-JD2, M13-JD3, M13-JD24, and M13-JD34 (Fig. 1) (19). There were three differences between our initial results and his. The GC at positions 5245 and 5246 (Fig. 2) was erroneously recorded as CG in our work; review of the gels shows that GC is correct. The sequence on one strand at positions 7213 and 7214 differed from that of the complementary strand, but the gel with the least band compression suggested that GC as found by Igarashi was correct.

Characterization of the KdpD protein, the sensor kinase of the K+‐translocating Kdp system of Escherichia coli

“…We found that the rate of expression of the KdpD protein is dependent on the K' concentration in the growth medium, leading to the detection of phosphorylation of the chromosome-encoded KdpD protein. In addition, we Kollmann and Altendorf, 1993Polarek et al, 1988Klionsky et al, 1984Yanish-Perron et al, 1985Puppe et al, 1992Walderhaug et al, 1992Walderhaug et al, 1992 this study Pharmacia this study Tabor and Richardson, 1985Tabor and Richardson, 1985Messing and Vieira, 1982 this study provide experimental evidence obtained by site-directed mutagenesis that the conserved histidine (His673) may represent the phosphorylation site in KdpD.…”

“…In plasmid pPV5, the kdpD gene is under the control of the tuc promoter. Plasmid pPV11, in which the kdpE gene is located downstream of the T7 promoter, was constructed by cloning the I-kb CluI-SmuI fragment of pPV2 (Walderhaug et al, 1992) into plasmid pT7-6 (Tabor and Richardson, 1985) digested with SmuI. Prior to that, the CluI ends of the 1-kb fragment had been filled-in using the Klenow fragment of DNA polymerase I of E. coli.…”

Truncation of Amino Acids 12–128 Causes Deregulation of the Phosphatase Activity of the Sensor Kinase KdpD of Escherichia coli

“…The kdpD gene has been cloned, sequenced, and overexpressed (12). The gene product KdpD is an integral protein of the cytoplasmic membrane (13).…”

“…(17), and pPV5-1 (21) were used. Plasmid pPV5-1 is a derivative of plasmid pPV5 (12), which contains five unique restriction sites that were introduced by silent mutation (21). All these plasmids are derivatives of pKK223-3, in which kdpD is under the control of the tac promoter.…”

The N-terminal Input Domain of the Sensor Kinase KdpD of Escherichia coli Stabilizes the Interaction between the Cognate Response Regulator KdpE and the Corresponding DNA-binding Site

“…Plasmid pBD/H673Q encodes the KdpD derivative with the inactivated phosphorylation site (KdpD/H673Q) (5). In plasmid pPV2, the kdpDE operon is under control of the lac promoter (1). Plasmids pPV2/1-395 and pPV2/1-395/D52N were obtained by digestion of plasmid pPV5-3/1-395 (11) with restriction enzymes NsiI and StuI, and the purified fragment was then ligated to similarly treated vector pPV2 or pPV2/D52N (see below), respectively.…”