title: 37-kDa laminin receptor precursor modulates cytotoxic necrotizing factor 1-mediated RhoA activation and bacterial uptake.
abstract: Cytotoxic necrotizing factor 1 (CNF1) is a bacterial toxin known to activate Rho GTPases and induce host cell cytoskeleton rearrangements. The constitutive activation of Rho GTPases by CNF1 is shown to enhance bacterial uptake in epithelial cells and human brain microvascular endothelial cells. However, it is unknown how exogenous CNF1 exhibits such phenotypes in eukaryotic cells. Here, we identified 37-kDa laminin receptor precursor (LRP) as the receptor for CNF1 from screening the cDNA library of human brain microvascular endothelial cells by the yeast two-hybrid system using the N-terminal domain of CNF1 as bait. CNF1-mediated RhoA activation and bacterial uptake were inhibited by exogenous LRP or LRP antisense oligodeoxynucleotides, whereas they were increased in LRP-overexpressing cells. These findings indicate that the CNF1 interaction with LRP is the initial step required for CNF1-mediated RhoA activation and bacterial uptake in eukaryotic cells.
authors.affiliation: Department of Pediatrics, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21287, USA.
authors.forename: Jin Woong
authors.initials: JW
authors.lastname: Chung
authors.forename: Suk Jin
authors.initials: SJ
authors.lastname: Hong
authors.forename: Kee Jun
authors.initials: KJ
authors.lastname: Kim
authors.forename: Daniel
authors.initials: D
authors.lastname: Goti
authors.forename: Monique F
authors.initials: MF
authors.lastname: Stins
authors.forename: Sooan
authors.initials: S
authors.lastname: Shin
authors.forename: Valina L
authors.initials: VL
authors.lastname: Dawson
authors.forename: Ted M
authors.initials: TM
authors.lastname: Dawson
authors.forename: Kwang Sik
authors.initials: KS
authors.lastname: Kim
mesh_terms: D001427:Bacterial Toxins; D002460:Cell Line; D003603:Cytotoxins / Q000502:physiology*; D004730:Endothelium, Vascular / Q000382:microbiology* / Q000502:physiology*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000502:physiology*; D004927:Escherichia coli Infections / Q000209:etiology / Q000378:metabolism; D029968:Escherichia coli Proteins; D016376:Oligonucleotides, Antisense; D011485:Protein Binding; D011498:Protein Precursors / Q000502:physiology*; D017474:Receptors, Laminin / Q000502:physiology*; D015398:Signal Transduction; D020742:rhoA GTP-Binding Protein / Q000502:physiology*
chemical_list: D001427:Bacterial Toxins; D003603:Cytotoxins; D029968:Escherichia coli Proteins; D016376:Oligonucleotides, Antisense; D011498:Protein Precursors; D017474:Receptors, Laminin; C060296:cytotoxic necrotizing factor type 1; D020742:rhoA GTP-Binding Protein