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Identification of Two Genes Encoding for the Late Acyltransferases of Lipid A in Klebsiella pneumoniae

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Abstract

Lipid A, the hydrophobic anchor of lipopolysaccharide, is an essential component in the outer membrane of most gram-negative bacteria. It is recognized by the TLR4/MD2 receptor of the innate immune system, which triggers an inflammatory response accompanied by massive overproduction of cytokines and leads to gram-negative septic shock. Human pathogen Klebsiella pneumoniae, which may synthesize two lipid A species, differs by the length of the secondary acyl chain. In this study, we identified two genes encoding the putative late acyltransferases of lipid A biosynthesis pathway in K. pneumoniae. Based on the sequence alignment, proteins YP_002239312.1 encoded by KPK3489 and YP_002239899.1 encoded by KPK4096 are homologous to E. coli LpxL, which were designated as LpxL1 and LpxL2, respectively. Functions of the two acyltransferases were confirmed by overexpressing the genes in E. coli, isolating lipid A and analyzing their structures using an ESI/MS. Like E. coli LpxL, K. pneumoniae LpxL1 transfers a C12:0 secondary acyl chain to the 2′-position of lipid A, while K. pneumoniae LpxL2 transfers a C14:0 secondary acyl chain to the 2′-position primary acyl chain of lipid A. These two acyltransferases might play important roles in the biosynthesis of lipid A and the innate immune system recognition in K. pneumoniae.

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References

  1. Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917

    Article  CAS  PubMed  Google Scholar 

  2. Brozek KA, Raetz CR (1990) Biosynthesis of lipid A in Escherichia coli. Acyl carrier protein-dependent incorporation of laurate and myristate. J Biol Chem 265:15410–15417

    CAS  PubMed  Google Scholar 

  3. Cai L, Li Y, Tao G, Guo W, Zhang C, Wang X (2013) Identification of three genes encoding for the late acyltransferases of lipid A in Cronobacter sakazakii. Mar drugs 11:377–386

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Clements A, Tull D, Jenney AW, Farn JL, Kim SH, Bishop RE, McPhee JB, Hancock RE, Hartland EL, Pearse MJ, Wijburg OL, Jackson DC, McConville MJ, Strugnell RA (2007) Secondary acylation of Klebsiella pneumoniae lipopolysaccharide contributes to sensitivity to antibacterial peptides. J Biol Chem 282:15569–15577

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Clementz T, Bednarski JJ, Raetz CR (1996) Function of the htrB high temperature requirement gene of Escherchia coli in the acylation of lipid A: htrB catalyzed incorporation of laurate. J Biol Chem 271:12095–12102

    Article  CAS  PubMed  Google Scholar 

  6. Lawlor MS, Hsu J, Rick PD, Miller VL (2005) Identification of Klebsiella pneumoniae virulence determinants using an intranasal infection model. Mol Microbiol 58:1054–1073

    Article  CAS  PubMed  Google Scholar 

  7. Li Y, Powell DA, Shaffer SA, Rasko DA, Pelletier MR, Leszyk JD, Scott AJ, Masoudi A, Goodlett DR, Wang X, Raetz CR, Ernst RK (2012) LPS remodeling is an evolved survival strategy for bacteria. Proc Natl Acad Sci USA 109:8716–8721

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Mey A, Ponard D, Colomb M, Normier G, Binz H, Revillard JP (1994) Acylation of the lipid A region of a Klebsiella pneumoniae LPS controls the alternative pathway activation of human complement. Mol Immunol 31:1239–1246

    Article  CAS  PubMed  Google Scholar 

  9. Miller SI, Ernst RK, Bader MW (2005) LPS, TLR4 and infectious disease diversity. Nat Rev Microbiol 3:36–46

    Article  CAS  PubMed  Google Scholar 

  10. Raetz CR, Garrett TA, Reynolds CM, Shaw WA, Moore JD, Smith DC Jr, Ribeiro AA, Murphy RC, Ulevitch RJ, Fearns C, Reichart D, Glass CK, Benner C, Subramaniam S, Harkewicz R, Bowers-Gentry RC, Buczynski MW, Cooper JA, Deems RA, Dennis EA (2006) Kdo2-lipid A of Escherichia coli, a defined endotoxin that activates macrophages via TLR-4. J Lipid Res 47:1097–1111

    Article  CAS  PubMed  Google Scholar 

  11. Raetz CR, Guan Z, Ingram BO, Six DA, Song F, Wang X, Zhao J (2009) Discovery of new biosynthetic pathways: the lipid A story. J Lipid Res 50(Suppl):S103–S108

    PubMed  PubMed Central  Google Scholar 

  12. Raetz CR, Reynolds CM, Trent MS, Bishop RE (2007) Lipid A modification systems in gram-negative bacteria. Annu Rev Biochem 76:295–329

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Raetz CR, Whitfield C (2002) Lipopolysaccharide endotoxins. Annu Rev Biochem 71:635–700

    Article  CAS  PubMed  Google Scholar 

  14. Sforza S, Silipo A, Molinaro A, Marchelli R, Parrilli M, Lanzetta R (2004) Determination of fatty acid positions in native lipid A by positive and negative electrospray ionization mass spectrometry. J Mass Spectrom 39:378–383

    Article  CAS  PubMed  Google Scholar 

  15. Six DA, Carty SM, Guan Z, Raetz CR (2008) Purification and mutagenesis of LpxL, the lauroyltransferase of Escherichia coli lipid A biosynthesis. Biochemistry 47:8623–8637

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Velkov T, Soon RL, Chong PL, Huang JX, Cooper MA, Azad MA, Baker MA, Thompson PE, Roberts K, Nation RL, Clements A, Strugnell RA, Li J (2013) Molecular basis for the increased polymyxin susceptibility of Klebsiella pneumoniae strains with under-acylated lipid A. Innate Immun 19:265–277

    Article  PubMed  Google Scholar 

  17. Vorachek-Warren MK, Ramirez S, Cotter RJ, Raetz CR (2002) A triple mutant of Escherichia coli lacking secondary acyl chains on lipid A. J Biol Chem 277:14194–14205

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (31300320), the Natural Science Foundation of Jiangsu Province (BK20130135), and Fundamental Research Funds for the Central Universities (JUSRP51411B).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, China

    Yanyan Li, Ye Li & Xiaoyuan Wang

  2. Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, China

    Yanyan Li, Jian Yun, Lu Liu, Ye Li & Xiaoyuan Wang

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  1. Yanyan Li
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  2. Jian Yun
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  3. Lu Liu
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Correspondence to Yanyan Li.

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Li, Y., Yun, J., Liu, L. et al. Identification of Two Genes Encoding for the Late Acyltransferases of Lipid A in Klebsiella pneumoniae . Curr Microbiol 73, 732–738 (2016). https://doi.org/10.1007/s00284-016-1117-6

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  • DOI: https://doi.org/10.1007/s00284-016-1117-6

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