Klebsiella pneumoniae type 3 fimbriae agglutinate yeast in a mannose-resistant manner

Stahlhut, Steen G.; Struve, Carsten; Krogfelt, Karen A.

doi:10.1099/jmm.0.036350-0

Cited by 21 publications

(16 citation statements)

References 35 publications

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“…Type 1 and type 3 fimbriae both bind to yeast surfaces [49]. A binding assay with mannan derived from Saccharomyces cerevisiae (the receptor compound) was performed to investigate the function of fimbriae.…”

Section: Resultsmentioning

confidence: 99%

“…pneumoniae strains express the type 1 and type 3 fimbriae [58–60]. Type 1 fimbrial expression is specifically detected by the mannose-sensitive agglutination of guinea pig RBCs, whereas type 3 fimbrial expression is detected by the agglutination of tannic acid-treated human erythrocytes in a mannose-resistant manner [17, 49]. This study is the first to describe the effects of the KP1_4563 gene on fimbriae.…”

Section: Discussionmentioning

confidence: 99%

“…Then, the minimum bacterial density (CFU/ml) required to agglutinate erythrocytes was measured. The expression of type 1 fimbriae was specifically detected via the mannose-sensitive agglutination of guinea pig red blood cells (RBCs) assays [49]. As described above, 2.5% guinea pig RBCs were mixed with a series of 2-fold dilutions of bacterial suspension with or without 0.25% mannose in 96-well, U-bottomed microtiter plates.…”

Section: Methodsmentioning

confidence: 99%

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The KP1_4563 gene is regulated by the cAMP receptor protein and controls type 3 fimbrial function in Klebsiella pneumoniae NTUH-K2044

Luo

Yang²,

et al. 2017

PLoS ONE

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Klebsiella pneumoniae (K. pneumoniae) is an opportunistic pathogen that can adhere to host cells or extracellular matrix via type 1 and type 3 fimbriae. KP1_4563 is a gene encoding a hypothetical protein in K. pneumoniae NTUH-K2044. KP1_4563 is located between the type 1 and type 3 fimbrial gene clusters and is likely associated with fimbrial function given its putative conserved domains of unknown function (DUF1471). Cyclic AMP receptor protein (CRP) regulates virulence-related gene expression and is a crucial transcriptional regulator in many bacteria. The predicted DNA recognition motif of CRP is present in the KP1_4563 promoter region. This study aimed to investigate the function of KP1_4563 in fimbriae and its transcriptional regulation mechanism by CRP. We generated Kp-Δ4563 mutant and complementation strains. We utilized phenotype and adhesion assays to evaluate the role of KP1_4563 in fimbriae. We conducted quantitative RT-PCR (qRT-PCR), LacZ fusion, electrophoretic mobility shift, and DNase I footprinting assays to study the transcriptional regulation of KP1_4563 gene by CRP. We found that KP1_4563 negatively regulates the function of type 3 fimbriae. Compared with NTUH-K2044, the absence of KP1_4563 enhanced the ability of Kp-Δ4563 to adhere to A549 cells. CRP negatively regulates KP1_4563 by directly binding to its promoter region. KP1_4563 plays an important role in type 3 fimbrial function. This novel insight will assist in the development of strategies for preventing K. pneumoniae infection.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The KP1_4563 gene is regulated by the cAMP receptor protein and controls type 3 fimbrial function in Klebsiella pneumoniae NTUH-K2044

Luo

Yang²,

et al. 2017

PLoS ONE

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“…mrkB, mrkC and mrkF encode the chaperone, usher and scaffolding proteins, respectively, which are responsible for fimbrial assembly/stabilization [65]. Type 3 fimbriae mediate in vitro adhesion to epithelial cells and kidney and lung tissues, most likely in a mannose-resistant manner [66]. Type 3 fimbriae act as a major contributor to K. pneumoniae biofilm formation, but play no REviEW Li, Zhao, Liu, Chen & Zhou future science group role in intestine and pulmonary infections [57].…”

Section: Fimbriaementioning

confidence: 99%

Molecular Pathogenesis of Klebsiella Pneumoniae

et al. 2014

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Typical Klebsiella pneumoniae is an opportunistic pathogen, which mostly affects those with weakened immune systems and tends to cause nosocomial infections. A subset of hypervirulent K. pneumoniae serotypes with elevated production of capsule polysaccharide can affect previously healthy persons and cause life-threatening community-acquired infections, such as pyogenic liver abscess, meningitis, necrotizing fasciitis, endophthalmitis and severe pneumonia. K. pneumoniae utilizes a variety of virulence factors, especially capsule polysaccharide, lipopolysaccharide, fimbriae, outer membrane proteins and determinants for iron acquisition and nitrogen source utilization, for survival and immune evasion during infection. This article aims to present the state-of-the-art understanding of the molecular pathogenesis of K. pneumoniae.

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“…By using antibodies against type 3 fimbriae, we found that that all recombinant strains expressed type 3 fimbriae at the same level, based on colony dot blot analysis (data not shown). Recently, we showed that type 3 fimbriae bind to the surface of yeast (53). We therefore utilized MrkD-expressing yeast strains to investigate the ability of the individual MrkD protein variant to bind yeast mannan (Fig.…”

Section: Figmentioning

confidence: 99%

Structural and Population Characterization of MrkD, the Adhesive Subunit of Type 3 Fimbriae

et al. 2013

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Type 3 fimbriae are adhesive organelles found in enterobacterial pathogens. The fimbriae promote biofilm formation on biotic and abiotic surfaces; however, the exact identity of the receptor for the type 3 fimbriae adhesin, MrkD, remains elusive. We analyzed naturally occurring structural and functional variabilities of the MrkD adhesin from Klebsiella pneumoniae and Escherichia coli isolates of diverse origins. We identified a total of 33 allelic variants of mrkD among 90 K. pneumoniae isolates and 10 allelic variants among 608 E. coli isolates, encoding 11 and 9 protein variants, respectively. Based on the level of accumulated silent variability between the alleles, mrkD was acquired a relatively long time ago in K. pneumoniae but recently in E. coli. However, unlike K. pneumoniae, mrkD in E. coli is actively evolving under a strong positive selection by accumulation of mutations, often targeting the same positions in the protein. Several naturally occurring MrkD protein variants from E. coli were found to be significantly less adherent when tested in a mannan-binding assay and showed reduced biofilm-forming capacity. Functional examination of the MrkD adhesin in flow chamber experiments determined that it interacts with Saccharomyces cerevisiae cells in a shear-dependent manner, i.e., the binding is catch-bond-like and enhanced under increasing shear conditions. Homology modeling strongly suggested that MrkD has a two-domain structure, comprising a pilin domain anchoring the adhesin to the fimbrial shaft and a lectin domain containing the binding pocket; this is similar to structures found in other catch-bond-forming fimbrial adhesins in enterobacteria.

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Klebsiella pneumoniae type 3 fimbriae agglutinate yeast in a mannose-resistant manner

Cited by 21 publications

References 35 publications

The KP1_4563 gene is regulated by the cAMP receptor protein and controls type 3 fimbrial function in Klebsiella pneumoniae NTUH-K2044

The KP1_4563 gene is regulated by the cAMP receptor protein and controls type 3 fimbrial function in Klebsiella pneumoniae NTUH-K2044

Molecular Pathogenesis of Klebsiella Pneumoniae

Structural and Population Characterization of MrkD, the Adhesive Subunit of Type 3 Fimbriae

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