The KP1_4563 Gene is Regulated by the CAMP Receptor Protein and Controls Type 3 Fimbrial Function in Klebsiella Pneumoniae NTUH-K2044

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 Jul 22

PMID 28732013

Citations 4

Authors

Mei Luo

Shiya Yang

Xuan Li

Pin Liu

Jian Xue

Xipeng Zhou

Kewen Su

Xuan Xu

Ying Qing

Jingfu Qiu

Yingli Li

Affiliations

Soon will be listed here.

Abstract

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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References

Langstraat J, Bohse M, Clegg S . Type 3 fimbrial shaft (MrkA) of Klebsiella pneumoniae, but not the fimbrial adhesin (MrkD), facilitates biofilm formation. Infect Immun. 2001; 69(9):5805-12. PMC: 98698. DOI: 10.1128/IAI.69.9.5805-5812.2001. View

Meyer M, Dimroth P, Bott M . Catabolite repression of the citrate fermentation genes in Klebsiella pneumoniae: evidence for involvement of the cyclic AMP receptor protein. J Bacteriol. 2001; 183(18):5248-56. PMC: 95405. DOI: 10.1128/JB.183.18.5248-5256.2001. View

Fic E, Bonarek P, Gorecki A, Kedracka-Krok S, Mikolajczak J, Polit A . cAMP receptor protein from escherichia coli as a model of signal transduction in proteins--a review. J Mol Microbiol Biotechnol. 2008; 17(1):1-11. DOI: 10.1159/000178014. View

Wu C, Huang Y, Fung C, Peng H . Regulation of the Klebsiella pneumoniae Kpc fimbriae by the site-specific recombinase KpcI. Microbiology (Reading). 2010; 156(Pt 7):1983-1992. DOI: 10.1099/mic.0.038158-0. View

Struve C, Bojer M, Krogfelt K . Identification of a conserved chromosomal region encoding Klebsiella pneumoniae type 1 and type 3 fimbriae and assessment of the role of fimbriae in pathogenicity. Infect Immun. 2009; 77(11):5016-24. PMC: 2772557. DOI: 10.1128/IAI.00585-09. View

Wilksch J, Yang J, Clements A, Gabbe J, Short K, Cao H . MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression. PLoS Pathog. 2011; 7(8):e1002204. PMC: 3161979. DOI: 10.1371/journal.ppat.1002204. View

Podschun R, Sievers D, Fischer A, Ullmann U . Serotypes, hemagglutinins, siderophore synthesis, and serum resistance of Klebsiella isolates causing human urinary tract infections. J Infect Dis. 1993; 168(6):1415-21. DOI: 10.1093/infdis/168.6.1415. View

Ou Q, Fan J, Duan D, Xu L, Wang J, Zhou D . Involvement of cAMP receptor protein in biofilm formation, fimbria production, capsular polysaccharide biosynthesis and lethality in mouse of Klebsiella pneumoniae serotype K1 causing pyogenic liver abscess. J Med Microbiol. 2016; 66(1):1-7. DOI: 10.1099/jmm.0.000391. View

Salazar J, Deng K, Tortorello M, Brandl M, Wang H, Zhang W . Genes ycfR, sirA and yigG contribute to the surface attachment of Salmonella enterica Typhimurium and Saintpaul to fresh produce. PLoS One. 2013; 8(2):e57272. PMC: 3579871. DOI: 10.1371/journal.pone.0057272. View

10.

Busby S, Ebright R . Transcription activation by catabolite activator protein (CAP). J Mol Biol. 1999; 293(2):199-213. DOI: 10.1006/jmbi.1999.3161. View

11.

Skorupski K, Taylor R . Cyclic AMP and its receptor protein negatively regulate the coordinate expression of cholera toxin and toxin-coregulated pilus in Vibrio cholerae. Proc Natl Acad Sci U S A. 1997; 94(1):265-70. PMC: 19310. DOI: 10.1073/pnas.94.1.265. View

12.

Favre-Bonte S, Darfeuille-Michaud A, Forestier C . Aggregative adherence of Klebsiella pneumoniae to human intestine-407 cells. Infect Immun. 1995; 63(4):1318-28. PMC: 173153. DOI: 10.1128/iai.63.4.1318-1328.1995. View

13.

Struve C, Bojer M, Krogfelt K . Characterization of Klebsiella pneumoniae type 1 fimbriae by detection of phase variation during colonization and infection and impact on virulence. Infect Immun. 2008; 76(9):4055-65. PMC: 2519443. DOI: 10.1128/IAI.00494-08. View

14.

Mukhopadhyay J, Sur R, Parrack P . Functional roles of the two cyclic AMP-dependent forms of cyclic AMP receptor protein from Escherichia coli. FEBS Lett. 1999; 453(1-2):215-8. DOI: 10.1016/s0014-5793(99)00719-x. View

15.

Zhang X, Garcia-Contreras R, Wood T . YcfR (BhsA) influences Escherichia coli biofilm formation through stress response and surface hydrophobicity. J Bacteriol. 2007; 189(8):3051-62. PMC: 1855844. DOI: 10.1128/JB.01832-06. View

16.

Ko W, Paterson D, Sagnimeni A, Hansen D, von Gottberg A, Mohapatra S . Community-acquired Klebsiella pneumoniae bacteremia: global differences in clinical patterns. Emerg Infect Dis. 2002; 8(2):160-6. PMC: 2732457. DOI: 10.3201/eid0802.010025. View

17.

Schmittgen T, Livak K . Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008; 3(6):1101-8. DOI: 10.1038/nprot.2008.73. View

18.

Eletsky A, Michalska K, Houliston S, Zhang Q, Daily M, Xu X . Structural and functional characterization of DUF1471 domains of Salmonella proteins SrfN, YdgH/SssB, and YahO. PLoS One. 2014; 9(7):e101787. PMC: 4092069. DOI: 10.1371/journal.pone.0101787. View

19.

Martinez-Antonio A, Collado-Vides J . Identifying global regulators in transcriptional regulatory networks in bacteria. Curr Opin Microbiol. 2003; 6(5):482-9. DOI: 10.1016/j.mib.2003.09.002. View

20.

Tarkkanen A, Virkola R, Clegg S, Korhonen T . Binding of the type 3 fimbriae of Klebsiella pneumoniae to human endothelial and urinary bladder cells. Infect Immun. 1997; 65(4):1546-9. PMC: 175168. DOI: 10.1128/iai.65.4.1546-1549.1997. View