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Chapter 20 : Genetic Analysis of the Escherichia coli K1 Capsule Gene Cluster

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Genetic Analysis of the Escherichia coli K1 Capsule Gene Cluster, Page 1 of 2

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Abstract:

Bacterial surface structures play a major role in the pathogenesis of infectious diseases. also continues to be the most common gram-negative organism that causes sepsis and meningitis in newborn infants. It is now known that produces more than 70 serologically distinct capsular antigens, or K antigens. Strains that produce the K1 capsule account for 80% of the isolates that cause neonatal meningitis and comprise the majority of isolates from neonates with septicemia without meningitis and acute pediatric pyelonephritis. The polysialic acid (PSA) capsule of K1 is essential for the virulence of the organism. The proteins necessary for the synthesis and expression of the K1 capsule are chromosomally encoded by the 17-kb kps gene cluster located near serA on the genetic map. Indeed, a major theme that has emerged from the study of capsule gene clusters in , , and is that the organization of capsule determinants in gram-negative bacteria is remarkably similar. The current model for the KpsMT transporter predicts that loops extending from KpsM (perhaps the CI loop) interact with the helical domain of KpsT, coupling the energy of ATP hydrolysis to the transport of PSA across the cytoplasmic membrane. The 17-kb kps gene cluster, which is divided into three functional regions, encodes the information necessary for polymer synthesis and expression at the cell surface.

Citation: Silver R. 1994. Genetic Analysis of the Escherichia coli K1 Capsule Gene Cluster, p 313-326. In Miller V, Kaper J, Portnoy D, Isberg R (ed), Molecular Genetics of Bacterial Pathogenesis. ASM Press, Washington, DC. doi: 10.1128/9781555818340.ch20

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Bacterial Cell Structure
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Integral Membrane Proteins
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Figures

Image of Figure 1
Figure 1

Genetic organization of the gene cluster of K1. The genes within the conserved regions 1 and 3 are designated while the genes within the K1-specific region 2 are designated Restriction endonuclease sites are indicated by the following abbreviations: B, II; C, I; H, dIII E, RI BH, HI P, I The arrows below the map indicate the direction of transcription for each region.

Citation: Silver R. 1994. Genetic Analysis of the Escherichia coli K1 Capsule Gene Cluster, p 313-326. In Miller V, Kaper J, Portnoy D, Isberg R (ed), Molecular Genetics of Bacterial Pathogenesis. ASM Press, Washington, DC. doi: 10.1128/9781555818340.ch20
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Image of Figure 2
Figure 2

Proposed transmembrane topology of KpsM (258 amino acids) in the cytoplasmic membrane ( ). Positions of the linker insertion mutations are indicated. Cells carrying the and mutations exhibited a null phenotype, while the and alleles did not affect protein function ( ).

Citation: Silver R. 1994. Genetic Analysis of the Escherichia coli K1 Capsule Gene Cluster, p 313-326. In Miller V, Kaper J, Portnoy D, Isberg R (ed), Molecular Genetics of Bacterial Pathogenesis. ASM Press, Washington, DC. doi: 10.1128/9781555818340.ch20
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Image of Figure 3
Figure 3

Proposed tertiary-structure model of the KpsT protein. The beta strands are represented by arrows, and the alpha helices are represented by cylinders. Each region of the protein is also indicated, and the locations of amino acid changes in mutant proteins are shown. Mutations were generated by hydroxylamine mutagenesis (shown boxed) and by site-directed (black circle) and saturation (black squares) mutagenesis by PCR ( ). Note the locations of the two cysteines in alpha helices 3 and 4 of the ATP-binding domain.

Citation: Silver R. 1994. Genetic Analysis of the Escherichia coli K1 Capsule Gene Cluster, p 313-326. In Miller V, Kaper J, Portnoy D, Isberg R (ed), Molecular Genetics of Bacterial Pathogenesis. ASM Press, Washington, DC. doi: 10.1128/9781555818340.ch20
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