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Chapter 15 : Regulation of Capsule Synthesis: Modification of the Two-Component Paradigm by an Accessory Unstable Regulator

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Regulation of Capsule Synthesis: Modification of the Two-Component Paradigm by an Accessory Unstable Regulator, Page 1 of 2

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Abstract:

Dramatic increases in expression correlate with increased capsule synthesis and generally can be ascribed to either of two control points for the regulatory system. Capsule synthesis under all these conditions is completely dependent on RcsB; RcsA appears to act as an accessory factor that allows modulation of RcsB activity. When high-level expression of fusions is selected at elevated temperatures, the primary location of the resulting mutations is in , the gene immediately clockwise to . Overproduction of RcsF increases capsule synthesis twofold, and mutations in decrease capsule synthesis two to threefold. Although there is no evidence for phosphorylation of RcsA, the temperature sensitivity of capsule synthesis is apparently best explained by temperature sensitivity of RcsA activity. The RcsB protein is relatively abundant in cells, and it is unclear whether there is any significant regulation of its activity during various growth conditions. RcsA is normally synthesized in very low amounts, and the protein is difficult to detect in wild-type cells, due to both low levels of synthesis and rapid degradation.

Citation: Gottesman S. 1995. Regulation of Capsule Synthesis: Modification of the Two-Component Paradigm by an Accessory Unstable Regulator, p 253-262. In Hoch J, Silhavy T (ed), Two-Component Signal Transduction. ASM Press, Washington, DC. doi: 10.1128/9781555818319.ch15

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Virulence Plasmid
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Klebsiella pneumoniae
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Figures

Image of FIGURE 1
FIGURE 1

Points of control for capsular polysaccharide synthesis. (A) Control pathway I: increased RcsB phosphorylation. In response to environmental signals or to changes in the cell surface as indicated in the box, the membrane protein sensor RcsC is activated, probably by a shift from phosphatase activity to kinase activity. The target of RcsC, the response regulator RcsB, is believed to act as a transcriptional activator when phosphorylated (shaded oval). High levels of activated RcsB allow transcription even when RcsA levels are low. Alternate kinase pathways (possibly involving RcsF) may also lead to RcsB activation. (See text for references and more detail.) (B) Control pathway II: increased accumulation of active RcsA. Increased capsule synthesis occurs when the amounts of active RcsA are increased. This can occur either when synthesis is increased, when degradation by Lon is blocked, or when RcsA is mutated (RcsA*) to improve its interaction with RcsB (shaded figure). RcsA activity is blocked at high temperatures. (C) Activation o f gene transcription. In the presence o f activated RcsB and RcsA, transcription occurs at high levels. High RcsB-P or high RcsA is sufficient to give high-level transcription. RcsA appears to be an auxiliary factor that stimulates RcsB activity. In this possible model for its action, we imagine cobinding of RcsA and RcsB to the promoter.

Citation: Gottesman S. 1995. Regulation of Capsule Synthesis: Modification of the Two-Component Paradigm by an Accessory Unstable Regulator, p 253-262. In Hoch J, Silhavy T (ed), Two-Component Signal Transduction. ASM Press, Washington, DC. doi: 10.1128/9781555818319.ch15
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Image of FIGURE 2
FIGURE 2

Conserved sequence elements in capsule regulatory proteins. (A) Domains of capsule regulators. The domains are shown to scale. The shaded regions bordering the periplasmic domain in RcsC are the hydrophobic regions believed to span the inner membrane. The kinase domain (solid) and effector domains (checkered) are defined as in ), and the DNA binding/activation domain (cross-hatched) as in ). The RmpA domain (hatched) is the region of high homology between RmpA and RmpA2, in addition to the homology seen in the DNA binding/activation domain ( ). (B) Alignment of DNA binding/activation domains from RcsB, RcsA, and RmpA proteins. The alignment between RcsA and RcsB is as in ), as is the consensus sequence. RmpA and RmpA2 were aligned with members of the conserved family using Blast ( ). The two positions marked with an asterisk are those expected to encode DNA binding site recognition residues, as predicted from mutagenesis studies on similar proteins and the position within the DNA binding helix of the putative helix-turn-helix ( ).

Citation: Gottesman S. 1995. Regulation of Capsule Synthesis: Modification of the Two-Component Paradigm by an Accessory Unstable Regulator, p 253-262. In Hoch J, Silhavy T (ed), Two-Component Signal Transduction. ASM Press, Washington, DC. doi: 10.1128/9781555818319.ch15
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References

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Tables

Generic image for table
TABLE 1

Genes affecting capsular polysaccharide synthesis in

Citation: Gottesman S. 1995. Regulation of Capsule Synthesis: Modification of the Two-Component Paradigm by an Accessory Unstable Regulator, p 253-262. In Hoch J, Silhavy T (ed), Two-Component Signal Transduction. ASM Press, Washington, DC. doi: 10.1128/9781555818319.ch15

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