Share This Article:

p54nrb, a PSF Protein Partner, Contributes to Meningitic Escherichia coli K1-Mediated Pathogenicities

Abstract Full-Text HTML Download Download as PDF (Size:787KB) PP. 1-10
DOI: 10.4236/ojapps.2012.21001    4,090 Downloads   9,087 Views  

ABSTRACT

IbeA is an important invasion determinant contributing to Escherichia coli K1 entry into brain microvascular endothelial cells (BMEC) that is a key step in the pathogenesis of E. coli meningitis. Our previous studies have shown that IbeA-induced signaling and E. coli K1 invasion is mediated by two IbeA-binding proteins, vimentin, which is constitutively present in the surface of human BMECs (HBMECs), and PSF, which is inducibly expressed in both mesenchymal (endothelium) and non-mesenchymal (epithelium) cells. However, it is unknown whether p54nrb, a PSF partner protein, could contribute to the pathogenesis of E. coli K1 meningitis. Here, we reported that a 54-kDa protein was identified by copurification with PSF through IbeA-affinity chromatography as an IbeA-binding protein, which is identical to p54nrb. Both p54nrb and PSF are RNA-binding proteins and share significant sequence homology. The specific interaction between IbeA and p54nrb was confirmed by Western blot and ligand overlay assays. Recombinant p54nrb blocked E. coli K1 invasion of human BMEC very effectively. Overexpressed p54nrb as a GFP fusion protein in the transfected 293T cells significantly enhanced E. coli K1 invasion. Furthermore, higher levels of surface p54nrb in the transfected 293T cells were detected by flow cytometry. These results suggest that the IbeA invasion protein of E. coli K1 interacts with p54nrb for bacterial invasion of human BMEC.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

L. He, F. Chi, T. Bo, L. Wang, C. Wu, A. Jong and S. Huang, "p54nrb, a PSF Protein Partner, Contributes to Meningitic Escherichia coli K1-Mediated Pathogenicities," Open Journal of Applied Sciences, Vol. 2 No. 1, 2012, pp. 1-10. doi: 10.4236/ojapps.2012.21001.

References

[1] S. H. Huang, C. A. Wass, Q. Fu, N. V. Prasadarao, M. F. Stins and K. S. Kim, “Escherichia coli Invasion of Brain Microvascular Endothelial Cells in Vitro and in Vivo: Molecular Cloning and Characterization of Invasion Gene ibe10,” Infection and Immunity, Vol. 63, No. 11, 1995, pp. 4470- 4475.
[2] S. H. Huang, M. Stins and K. S. Kim, “Bacterial Penetration across the Blood-Brain Barrier during the Development of Neonatal Meningitis,” Microbes and Infection, Vol. 2, No. 10, 2000, pp. 1237-1244. doi:10.1016/S1286-4579(00)01277-6
[3] S. H. Huang and A. Jong, “Cellular Mechanisms of Micro- bial Proteins Contributing to Invasion of the Blood-Brain Barrier,” Cellular Microbiology, Vol. 3, No. 5, 2001, pp. 277-287. doi:10.1046/j.1462-5822.2001.00116.x
[4] K. S. Kim, “Pathogenesis of Bacterial Meningitis: From Bacteraemia to Neuronal Injury,” Nature Reviews Neuroscience, Vol. 4, No. 5, 2003, pp. 376-385. doi:10.1038/nrn1103
[5] S. H. Huang, Z. S. Wan, Y. H. Chen, A. Y. Jong and K. S. Kim, “Further Characterization of Escherichia coli Brain Microvascular Endothelial Cell Invasion Gene ibeA by Deletion, Complementation, and Protein Expression,” Journal of Infectious Diseases, Vol. 183, No. 7, 2001, pp. 1071-1078. doi:10.1086/319290
[6] S. H. Huang, Y. H. Chen, G. Y. Kong, S. H. Chen, M. Borodovsky, J. Besemer and A. Y. Jong, “A Novel Genetic Island of Meningitic E. coli K1 Containing the ibeA Invasion Gene (GimA): Functional Annotation and Carbon Source-Regulated Invasion of Human Endothelial Cells,” Functional & Integrative Genomics, Vol. 1, No. 5, 2001, pp. 312-322. doi:10.1007/s101420100039
[7] S. H. Huang, P. Germon and A. Y. Jong, “The Invasion- Associated Genetic Island GimA in Meningitic E. coli K1,” In: F. Columbus, Ed., Focus on Meningitis Research, Nova Science Publisher, New York, 2004, pp. 97-114.
[8] S. Bonacorsi, O. Clermont, V. Houdouin, C. Cordevant, N. Brahimi, A. Marecat, C. Tinsley, X. Nassif, M. Lange and E. Bingen, “Molecular Analysis and Experimental Virulence of French and North American Escherichia coli Neonatal Meningitis Isolates: Identification of a New Virulent Clone,” Journal of Infectious Diseases, Vol. 188, No. 7, 2003, pp. 1895-1906. doi:10.1086/375347
[9] Y. H. Chen, S. H. Chen, A. Jong, Z. Y. Zhou, W. Li, K. Suzuki and S. H. Huang, “Enhanced Escherichia coli Invasion of Human Brain Microvascular Endothelial Cells Is Associated with Alterations in Cytoskeleton Induced by Nicotine,” Cellular Microbiology, Vol. 4, No. 8, 2002, pp. 503-514. doi:10.1046/j.1462-5822.2002.00209.x
[10] F. Chi, T. Jong, L. Wang, Y. Ouyang, C. Wu, W. Li and S. H. Huang, “Vimentin-Mediated Signaling Is Required for IbeA+ E. coli K1 Invasion of Human Brain Microvascular Endothelial Cells,” Biochemical Journal, Vol. 427, No. 1, 2010, pp. 79-90. doi:10.1042/BJ20091097
[11] F. Chi, L. Wang, X. Zheng, A. Jong and S. H. Huang, “Recruitment of α7 Nicotinic Acetylcholine Receptor to Caveolin-1-enriched Lipid Rafts is Required for Nicotine-Enhanced Escherichia coli K1 Entry into Brain Endothelial Cells,” Future Microbiology, Vol. 6, No. 8, 2011, PP. 953-966.
[12] T. Kihara, S. Shimohama, H. Sawada, K. Honda, T. Nakamizo and H. Shibasaki, “Alpha 7 Nicotinic Receptor Transduces Signals to Phosphatidylinositol 3-Kinase to Block A Beta-Amyloid-Induced Neurotoxicity,” Journal of Biological Chemistry, Vol. 276, No. 17, 2001, pp. 13541-13546.
[13] Y. Zou, L. He and S. H. Huang, “Identification of a Surface Protein on Human Brain Microvascular Endothelial Cells as Vimentin Interacting with Escherichia coli Invasion Protein IbeA,” Biochemical and Biophysical Research Communications, Vol. 351, No. 3, 2006, pp. 625-630. doi:10.1016/j.bbrc.2006.10.091
[14] N. V. Prasadarao, C. A. Wass, S. H. Huang and K. S. Kim, “Identification and Characterization of a Novel Ibe10 Binding Protein that Contributes to Escherichia coli Invasion of Brain Microvascular Endothelial Cells,” Infection and Immunity, Vol. 67, No. 3, 1999, pp. 1131-1138.
[15] Y. Zou, L. He, C. H. Wu, H. Cao, Z. H. Xie, Y. Wang, A. Y. Jong and S. H. Huang, “PSF Is an IbeA-Binding Protein Contributing to Meningitic Escherichia coli K1 Invasion of Human Brain Microvascular Endothelial Cells,” Medical Microbiology and Immunology, Vol. 196, No. 3, 2007, pp. 135-143. doi:10.1007/s00430-006-0034-x
[16] C. L. Bladen, D. Udayakumar, Y. Takeda and W. S. Dynan, “Identification of the Polypyrimidine Tract Binding Protein-Associated Splicing Factor p54(nrb) Complex as a Candidate DNA Double-Strand Break Rejoining Factor,” Journal of Biological Chemistry, Vol. 280, No. 7, 2005, pp. 5205-5210. doi:10.1074/jbc.M412758200
[17] J. F. Sinclair and A. D. O'Brien, “Cell Surface-Localized Nucleolin Is a Eukaryotic Receptor for the Adhesin Intimin-Gamma of Enterohemorrhagic Escherichia coli O157:H7,” Journal of Biological Chemistry, Vol. 277, No. 4, 2002, pp. 2876-2885. doi:10.1074/jbc.M110230200
[18] S. F. Altschul, T. L. Madden, A. A. Sch?ffer, J. Zhang, Z. Zhang, W. Miller and D. J. Lipman, “Gapped BLAST and PSI-BLAST: A New Generation of Protein Database Search Programs,” Nucleic Acids Research, Vol. 25, No. 7, 1997, pp. 3389-3402. doi:10.1093/nar/25.17.3389
[19] K. Guan and J. E. Dixon, “Eukaryotic Proteins Expressed in Escherichia coli: An Improved Thrombin Cleavage and Purification Procedure of Fusion Proteins with Glutathione S-Transferase,” Analytical Biochemistry, Vol. 192, No. 2, 1991, pp. 262-267. doi:10.1016/0003-2697(91)90534-Z
[20] R. Peng, B. T. Dye, I. Perez, D. C. Barnard, A. B. Thom- pson and J. G. Patton, “PSF and p54nrb Bind a Conserved Stem in U5 snRNA,” RNA, Vol. 8, No. 10, 2002, pp. 1334-1347. doi:10.1017/S1355838202022070
[21] B. T. Dye and J. G. Patton, “An RNA Recognition Motif (RRM) Is Required for the Localization of PTB Associated Splicing Factor (PSF) to Subnuclear Speckles,” Experimental Cell Research, Vol. 263, No. 1, 2001, pp. 131- 144. doi:10.1006/excr.2000.5097
[22] L. Braun, B. Ghebrehiwet and P. Cossart, “gC1q-R/p32, a C1q-Binding Protein, Is a Receptor for the InIB Invasion Protein of Listeria monocytogenes,” EMBO Journal, Vol. 19, No. 7, 2000, pp. 1458-1466. doi:10.1093/emboj/19.7.1458
[23] R. E. Kingston, C. A. Chen and H. Okayama, “Calcium Phosphate Transfection,” In: J. S. Bonifacino, M. Dasso, J. B. Harford, J. L. Schwartz and K. M. Yamada, Eds., Current Protocols in Cell Biology, John Wiley & Sons, New York, 2003. doi:10.1002/0471143030.cb2003s19
[24] S. Watt, P. Lanotte, L. Mereghetti, M. M. Schouleur, B. Picard and R. Quentin, “Escherichia coli Strains from Pregnant Women and Neonates: Intraspecies Genetic Distribution and Prevalence of Virulence Factors,” Journal of Clinical Microbiology, Vol. 41, No. 5, 2003, pp. 1929- 1935. doi:10.1128/JCM.41.5.1929-1935.2003
[25] N. V. Prasadarao, P. K. Srivastava, R. S. Rudrabhatla, K. S. Kim, S. H. Huang and S. K. Sukumaran, “Cloning and Expression of the Escherichia coli K1 Outer Membrane Protein A Receptor, a gp96 Homologue,” Infection and Immunity, Vol. 71, No. 4, 2003, pp. 1680-1688. doi:10.1128/IAI.71.4.1680-1688.2003
[26] K. J. Kim, J. W. Chung and K. S. Kim, “67-kDa Laminin Receptor Promotes Internalization of Cytotoxic Necrotizing Factor 1-Expressing Escherichia coli K1 into Human Brain Microvascular Endothelial Cells,” Journal of Biological Chemistry, Vol. 280, No. 2, 2005, pp. 1360-1368. doi:10.1074/jbc.M410176200
[27] A. H. Fox, C. S. Bond and A. I. Lamond, “P54nrb Forms a Heterodimer with PSP1 That Localizes to Paraspeckles in an RNA-Dependent Manner,” Molecular Biology of the Cell, Vol. 16, No. 11, 2005, pp. 5304-5315. doi:10.1091/mbc.E05-06-0587
[28] L. Liu, N. Xie, P. Rennie, J. R. Challis, M. Gleave, S. J. Lye and X. Dong, “Consensus PP1 Binding Motifs Regulate Transcriptional Corepression and Alternative RNA Splicing Activities of the Steroid Receptor Coregulators, p54nrb and PSF,” Molecular Endocrinology, Vol. 25, No. 7, 2011, pp. 1197-1210. doi:10.1210/me.2010-0517
[29] K. Thiele, D. Riemann, S. A. Navarrete, J. Langner and A Kehlen, “Cell-Cell Contact of Human T Cells with Fibroblasts Changes Lymphocytic mRNA Expression: Increased mRNA Expression of Interleukin-17 and Interleukin-17 Receptor,” European Cytokine Network, Vol. 11, No. 1, 2000, pp. 53-58.
[30] K. Ishitani, T. Yoshida, H. Kitagawa, H. Ohta, S. Nozawa and S. Kato, “p54nrb Acts as a Transcriptional Coactivator for Activation Function 1 of the Human Androgen Receptor,” Biochemical and Biophysical Research Communications, Vol. 306, No. 3, 2003, pp. 660-665. doi:10.1016/S0006-291X(03)01021-0
[31] O. Adegbola and G. R. Pasternack, “A pp32-Retinoblas- toma Protein Complex Modulates Androgen Receptor-Mediated Transcription and Associates with Components of the Splicing Machinery,” Biochemical and Biophysical Research Communications, Vol. 334, No. 2, 2005, pp. 702-708. doi:10.1016/j.bbrc.2005.06.153
[32] A. Proteau, S. Blier, A. L. Albert, S. B. Lavoie, A. M. Traish and M. Vincent, “The Multifunctional Nuclear Protein p54nrb Is Multiphosphorylated in Mitosis and Interacts with the Mitotic Regulator Pin1,” Journal of Molecular Biology, Vol. 436, No. 4, 2005, pp. 1163-1172. doi:10.1016/j.jmb.2004.12.034
[33] T. Adam, M. Arpin, M. C. Prevost, P. Gounon and P. J. Sansonetti, “Cytoskeletal Rearrangements and the Functional Role of T-Plastin during Entry of Shigella flexneri into HeLa Cells,” Journal of Cell Biology, Vol. 192, No. 2, 1995, pp. 367-381. doi:10.1083/jcb.129.2.367
[34] D. Zhou, M. S. Mooseker and J. E. Galan, “An Invasion-Associated Salmonella Protein Modulates the Actin-Bundling Activity of Plastin,” Proceeding of the National Academy of Sciences of the United States of America, Vol. 96, No. 18, 1999, pp. 10176-10181.
[35] N. Manel, F. J. Kim, S. Kinet, N. Taylor, M. Sitbon and J. L. Battini, “The Ubiquitous Glucose Transporter GLUT-1 Is a Receptor for HTLV,” Cell, Vol. 115, No. 4, 2003, pp. 449-459. doi:10.1016/S0092-8674(03)00881-X
[36] M. Srivastava and H. B. Pollard, “Molecular Dissection of Nucleolin’s Role in Growth and Cell Proliferation: New Insights,” FASEB Journal, Vol. 13, No. 14, 1999, pp. 1911-1922.
[37] F. Chi, T. Bo, C. H. Wu, A. Jong and S. H. Huang, “Vimentin and PSF Contribute Cooperatively to IbeA+ Escherichia coli K1-Induced Activation and Nuclear Translocation of NF-κB in Human Brain Microvascular Endothelial Cells,” PLoS One, Accepted Pending Minor Revision.
[38] X. Che, F. Chi, L. Wang, T. Jong, C. H. Wu, X. Wang and S. H. Huang, “Involvement of IbeA in Meningitic Escherichia coli K1-Induced Polymorphonuclear Leukocyte Transmigration across Brain Endothelial Cells,” Brain Pathology, Vol. 21, No. 4, 2011, pp. 389-404. doi:10.1111/j.1750-3639.2010.00463.x

  
comments powered by Disqus

Copyright © 2018 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.