The Chlamydia pneumoniae Invasin Protein Pmp21 Recruits the EGF Receptor for Host Cell Entry

Infection of mammalian cells by the strictly intracellular pathogens Chlamydiae requires adhesion and internalization of the infectious Elementary Bodies (EBs). The components of the latter step were unknown. Here, we identify Chlamydia pneumoniae Pmp21 as an invasin and EGFR as its receptor. Modulation of EGFR surface expression evokes correlated changes in EB adhesion, internalization and infectivity. Ectopic expression of EGFR in EGFR-negative hamster cells leads to binding of Pmp21 beads and EBs, thus boosting the infection. EB/Pmp21 binding and invasion of epithelial cells results in activation of EGFR, recruitment of adaptors Grb2 and c-Cbl and activation of ERK1/2, while inhibition of EGFR or MEK kinase activity abrogates EB entry, but not attachment. Binding of Grb2 and c-Cbl by EGFR is essential for infection. This is the first report of an invasin-receptor interaction involved in host-cell invasion by any chlamydial species.

[1]  B. Coombes,et al.  Identification of MEK‐ and phosphoinositide 3‐kinase‐dependent signalling as essential events during Chlamydia pneumoniae invasion of HEp2 cells , 2002, Cellular microbiology.

[2]  L. Norkin,et al.  Lipid rafts, caveolae, caveolin-1, and entry by Chlamydiae into host cells. , 2003, Experimental cell research.

[3]  E. Wolf,et al.  The epidermal growth factor receptor ligands at a glance , 2009, Journal of cellular physiology.

[4]  W R Taylor,et al.  On the tertiary structure of the extracellular domains of the epidermal growth factor and insulin receptors. , 1987, Biochimica et biophysica acta.

[5]  K. Haglund,et al.  The role of ubiquitylation in receptor endocytosis and endosomal sorting , 2012, Journal of Cell Science.

[6]  T. Hackstadt,et al.  Chlamydia trachomatis Induces Remodeling of the Actin Cytoskeleton during Attachment and Entry into HeLa Cells , 2002, Infection and Immunity.

[7]  T. Hackstadt,et al.  A chlamydial type III translocated protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[8]  J. Hegemann,et al.  Members of the Pmp protein family of Chlamydia pneumoniae mediate adhesion to human cells via short repetitive peptide motifs , 2010, Molecular microbiology.

[9]  S. Gygi,et al.  Multiple mechanisms collectively regulate clathrin-mediated endocytosis of the epidermal growth factor receptor , 2010, The Journal of cell biology.

[10]  L. Campbell,et al.  Chlamydia pneumoniae — an infectious risk factor for atherosclerosis? , 2004, Nature Reviews Microbiology.

[11]  R. Fouchier,et al.  The Epidermal Growth Factor Receptor (EGFR) Promotes Uptake of Influenza A Viruses (IAV) into Host Cells , 2010, PLoS pathogens.

[12]  K. Gevaert,et al.  The expression, processing and localization of polymorphic membrane proteins in Chlamydia pneumoniae strain CWL029 , 2002, BMC Microbiology.

[13]  M. Hammerschlag,et al.  Use of HEp-2 cells for improved isolation and passage of Chlamydia pneumoniae , 1992, Journal of clinical microbiology.

[14]  T. Compton,et al.  Epidermal Growth Factor Receptor Is Not Required for Human Cytomegalovirus Entry or Signaling , 2007, Journal of Virology.

[15]  J. Hegemann,et al.  Chlamydial Adhesion and Adhesins , 2012 .

[16]  J. Schlessinger Cell Signaling by Receptor Tyrosine Kinases , 2000, Cell.

[17]  Walter Kolch,et al.  Functional proteomics to dissect tyrosine kinase signalling pathways in cancer , 2010, Nature Reviews Cancer.

[18]  G. Zhong,et al.  Activation of Raf/MEK/ERK/cPLA2 Signaling Pathway Is Essential for Chlamydial Acquisition of Host Glycerophospholipids* , 2004, Journal of Biological Chemistry.

[19]  J. Massagué,et al.  A single mutation affects both N-acetylglucosaminyltransferase and glucuronosyltransferase activities in a Chinese hamster ovary cell mutant defective in heparan sulfate biosynthesis. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[20]  A. Sorkin,et al.  Endocytosis and signalling: intertwining molecular networks , 2009, Nature Reviews Molecular Cell Biology.

[21]  P. Jeffrey,et al.  Structural basis for inhibition of the epidermal growth factor receptor by cetuximab. , 2005, Cancer cell.

[22]  R. Lahesmaa,et al.  Chlamydia pneumoniae entry into epithelial cells by clathrin-independent endocytosis. , 2012, Microbial pathogenesis.

[23]  S. Huong,et al.  Epidermal growth factor receptor is a cellular receptor for human cytomegalovirus , 2003, Nature.

[24]  Y. Kido,et al.  Requirement of autophosphorylated tyrosine 992 of EGF receptor and its docking protein phospholipase Cγ1 for membrane ruffle formation , 2003, FEBS letters.

[25]  M. Jaramillo,et al.  Effect of the anti-receptor ligand-blocking 225 monoclonal antibody on EGF receptor endocytosis and sorting. , 2006, Experimental cell research.

[26]  J. Esko,et al.  Animal cell mutants defective in glycosaminoglycan biosynthesis. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[27]  A. Sorkin Internalization of the epidermal growth factor receptor: role in signalling. , 2001, Biochemical Society transactions.

[28]  A. Dautry‐Varsat,et al.  Chlamydia infection of epithelial cells expressing dynamin and Eps15 mutants: clathrin-independent entry into cells and dynamin-dependent productive growth. , 1999, Journal of cell science.

[29]  Epidermal Growth Factor Receptor Dimerization and Activation Require Ligand-Induced Conformational Changes in the Dimer Interface , 2005, Molecular and Cellular Biology.

[30]  R. Iozzo,et al.  Decorin Binds to a Narrow Region of the Epidermal Growth Factor (EGF) Receptor, Partially Overlapping but Distinct from the EGF-binding Epitope* , 2002, The Journal of Biological Chemistry.

[31]  D. Kalman,et al.  RNA Interference Screen Identifies Abl Kinase and PDGFR Signaling in Chlamydia trachomatis Entry , 2008, PLoS pathogens.

[32]  T. Hackstadt,et al.  Recent insights into the mechanisms of Chlamydia entry , 2005, Cellular microbiology.

[33]  K. Hybiske,et al.  Mechanisms of Chlamydia trachomatis Entry into Nonphagocytic Cells , 2007, Infection and Immunity.

[34]  R. Seger,et al.  Detection of partially phosphorylated forms of ERK by monoclonal antibodies reveals spatial regulation of ERK activity by phosphatases , 2000, FEBS letters.

[35]  J. Hegemann,et al.  Antigenic and molecular analyses of different Chlamydia pneumoniae strains , 1997, Journal of clinical microbiology.

[36]  C. Elwell,et al.  Chlamydia trachomatis Co-opts the FGF2 Signaling Pathway to Enhance Infection , 2011, PLoS pathogens.

[37]  A. Sorkin,et al.  Endocytosis and intracellular trafficking of ErbBs. , 2008, Experimental cell research.

[38]  J. Hegemann,et al.  Chlamydia pneumoniae GroEL1 Protein Is Cell Surface Associated and Required for Infection of HEp-2 Cells , 2008, Journal of bacteriology.

[39]  A. Wells EGF receptor. , 1999, The international journal of biochemistry & cell biology.

[40]  Xuejun Jiang,et al.  Grb2 regulates internalization of EGF receptors through clathrin-coated pits. , 2003, Molecular biology of the cell.

[41]  T. Meyer,et al.  From the inside out – processing of the Chlamydial autotransporter PmpD and its role in bacterial adhesion and activation of human host cells , 2004, Molecular microbiology.

[42]  John Kuriyan,et al.  An Allosteric Mechanism for Activation of the Kinase Domain of Epidermal Growth Factor Receptor , 2006, Cell.

[43]  Edouard C. Nice,et al.  Crystal Structure of a Truncated Epidermal Growth Factor Receptor Extracellular Domain Bound to Transforming Growth Factor α , 2002, Cell.