title: Internalization of Escherichia coli by human renal epithelial cells is associated with tyrosine phosphorylation of specific host cell proteins.
abstract: Human renal epithelial cells are capable of internalizing Escherichia coli regardless of whether the bacteria are isolated from individuals with pyelonephritis or from healthy volunteers. In this study, we investigated the role of host cell tyrosine kinase activity in internalization. We found that internalization of both fecal and pyelonephritis isolates is blocked by tyrosine kinase inhibitors. We found increased intensity of two tyrosine-phosphorylated proteins, with relative mobilities of approximately 123,000 and 110,000, in Western blots of extracts from human renal epithelial cells infected with E. coli. The increased intensity of these tyrosine-phosphorylated proteins was observed only in the Triton X-100-insoluble fraction, suggesting that these proteins could be associated with the cytoskeleton. Increased tyrosine phosphorylation of these proteins upon E. coli infection was observed in both transformed and primary human renal epithelial cells and in cells infected with several different strains of E. coli isolated from the feces of healthy individuals or from the blood or urine of patients with pyelonephritis. The increased tyrosine phosphorylation of these proteins required live bacteria and was blocked by tyrosine kinase inhibition but not by protein synthesis inhibitors or cytochalasin D. These experiments establish a strong link between E. coli internalization and host cell signaling through tyrosine kinases in human kidney cells and provide evidence that specific proteins are involved in these processes.
authors.affiliation: Department of Medicine, University of Maryland School of Medicine, Baltimore 21201, USA.
authors.forename: L M
authors.initials: LM
authors.lastname: Palmer
authors.forename: T J
authors.initials: TJ
authors.lastname: Reilly
authors.forename: S J
authors.initials: SJ
authors.lastname: Utsalo
authors.forename: M S
authors.initials: MS
authors.lastname: Donnenberg
mesh_terms: D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D004848:Epithelium / Q000382:microbiology; D004926:Escherichia coli / Q000502:physiology*; D004927:Escherichia coli Infections / Q000201:enzymology; D006801:Humans; D007668:Kidney / Q000382:microbiology*; D007674:Kidney Diseases / Q000201:enzymology; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011704:Pyelonephritis / Q000382:microbiology; D015398:Signal Transduction / Q000502:physiology; D014443:Tyrosine / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases