umu.sePublikasjoner
Endre søk
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
YopH dephosphorylates Cas and Fyn-binding protein in macrophages
Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten).
Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten).
Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten).
Vise andre og tillknytning
1999 (engelsk)Inngår i: Microbial Pathogenesis, ISSN 0882-4010, E-ISSN 1096-1208, Vol. 27, nr 4, s. 231-242Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

The tyrosine phosphatase YopH is an essential virulence effector of pathogenic Yersinia spp. YopH, which is translocated from extracellularly located bacteria into interacting target cells, blocks phagocytosis by professional phagocytes. We show here that immunoprecipitation of YopH from lysates of J774 cells infected with Y. pseudotuberculosis expressing an inactive form of YopH resulted in co-precipitation of certain phosphotyrosine proteins. The association between the inactive YopH and phosphotyrosine proteins in the 120 kDa range was rapid and could be detected after 2 min of infection. The proteins were identified as the docking proteins Cas and Fyn-binding protein (FYB). Upon infection of J774 cells with Y. pseudotuberculosis lacking YopH expression both of these proteins became tyrosine phosphorylated. Moreover, this infection caused recruitment of Cas to peripheral focal complexes, and FYB was relocalized to areas surrounding these structures. Both Cas and FYB became dephosphorylated upon infection with Y. pseudotuberculosis expressing active YopH, and this was associated with disruption of focal complexes. With regard to the previous identification of Cas and focal complexes as targets of YopH in HeLa cells, the present study supports an important role for these targets in a general mechanism of bacterial uptake. 

sted, utgiver, år, opplag, sider
Academic Press, 1999. Vol. 27, nr 4, s. 231-242
Emneord [en]
Yersinia pseudotuberculosis, phagocytosis, Cas, FYB, focal complexes, PTPase
HSV kategori
Identifikatorer
URN: urn:nbn:se:umu:diva-3442DOI: 10.1006/mpat.1999.0301ISI: 000083093100006PubMedID: 10502464OAI: oai:DiVA.org:umu-3442DiVA, id: diva2:142137
Tilgjengelig fra: 2004-01-30 Laget: 2004-01-30 Sist oppdatert: 2019-01-24bibliografisk kontrollert
Inngår i avhandling
1. Effects of invasin and YopH of Yersinia pseudotuberculosis on host cell signaling
Åpne denne publikasjonen i ny fane eller vindu >>Effects of invasin and YopH of Yersinia pseudotuberculosis on host cell signaling
2004 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Alternativ tittel[sv]
Effekter av proteinerna invasin och YopH från bakterien Yersinia pseudotuberculosis på värdcellen
Abstract [en]

Integrins are a large family of membrane-spanning heterodimeric (αβ) receptors that bind to ligands on other cells or to extracellular matrix (ECM) proteins. These receptors mediate bidirectional signaling over the cell membrane to induce signaling cascades mediating functions as cell adhesion, spreading and migration. This signaling takes place at cell-matrix adhesions, which are sites where clustered and ligand-bound integrins connect to and mediate stabilization of the actin cytoskeleton, and induce signaling cascades. Integrins have a short cytoplasmic tail that is crucial for the bidirectional signaling, and the β1-integrin subunit exists in five splice variants only differing in the membrane-distal part of the cytoplasmic tail. This region of the almost ubiquitously expressed β1-integrin, β1A, contains two protein tyrosine motifs (NPXYs) interspaced with a threonine-rich region, while this region of the β1B splice variant is completely different and lacks known motifs. In contrast to the β1A-integrin, the β1B variant cannot mediate cell-matrix adhesion formation following binding to ECM ligands.

The enteropathogenic bacterium Yersinia pseudotuberculosis binds to β1-integrins on the host cell with invasin, and this stimulates uptake of the bacterium. However, upon binding to the host cell, pathogenic Yersinia strains inject virulence effectors that block uptake. One effector responsible for the blocking is a tyrosine phosphatase, YopH. We identified the targets for this effector in the macrophage-like cell line J774A.1, which represent a professional phagocyte and thus is the likely target cell for the antiphagocytic effect of Yersinia. Two YopH target proteins were p130Cas and ADAP, of which the latter interestingly is an adapter protein specifically expressed in hematopoietic cells. ADAP has previously been implicated to participate in Fc-receptor-mediated phagocytosis and in communication between T-cell receptors and integrins.

We also studied the importance of the cytoplasmic tail of β1-integrin for uptake of Yersinia. The GD25 cell line, which is a fibroblast-like cell line that lacks endogenous β1-integrins, was used together with GD25 cells transfected with β1B, β1Α or cytoplasmic tail mutants of β1A. These studies revealed that β1B-integrins could bind to invasin but not mediate uptake of Yersinia, while β1A both bound to invasin and mediated uptake. The first NPXY motif (unphosphorylated) and the double-threonines of the unique part of β1A were important for the ability of integrin to mediate uptake of Yersinia. These studies lead to the interesting finding that, when these cells were allowed to spread on invasin, those that expressed β1A spread as normal fibroblasts while for β1B-integrin-expressing cells, only finger-like protrusions of filopodia were formed. This provided us with a tool to study formation of filopodia without interference of the tightly linked process of lamellipodia formation. Initially, proteins that localized to the tip complex of these filopodia were identified. These were talin, VASP and interestingly the p130Cas-Crk-DOCK180 scaffold, while FAK, paxillin and vinculin were absent. In addition, VASP, p130Cas and Crk were shown to be important for the filopodia formation in GD25β1B. Further, the role of the actin motor myosin X, which previously has been implicated in formation of filopodia, was studied in the GD25Β1B cells and it was shown that myosin X not was important for filopodia formation, but that it recruited FAK and vinculin to the tip complexes of filopodia.

sted, utgiver, år, opplag, sider
Umeå: Molekylärbiologi (Teknisk-naturvetenskaplig fakultet), 2004. s. 67
Emneord
Molecular biology, β1-integrin, Yersinia pseudotuberculosis, Invasin, YopH, Cell-Matrix Adhesion, Filopodia, Cell Spreading, Molekylärbiologi
HSV kategori
Forskningsprogram
molekylärbiologi
Identifikatorer
urn:nbn:se:umu:diva-183 (URN)91-7305-588-3 (ISBN)
Disputas
2004-03-05, Major Groove, 6L, NUS, Inst f molekylärbiologi, Umeå Universitet, 901 87 Umeå, Umeå, 09:00 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2004-01-30 Laget: 2004-01-30 Sist oppdatert: 2019-01-22bibliografisk kontrollert
2. Interactions between Yersinia pseudotuberculosis and host cells - role of invasin and YopH
Åpne denne publikasjonen i ny fane eller vindu >>Interactions between Yersinia pseudotuberculosis and host cells - role of invasin and YopH
1999 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
sted, utgiver, år, opplag, sider
Umeå: Umeå universitet, 1999. s. 72
Serie
Umeå University medical dissertations, ISSN 0346-6612 ; 634
HSV kategori
Identifikatorer
urn:nbn:se:umu:diva-65079 (URN)91-7191-732-2 (ISBN)
Disputas
Föreläsningssalen, Institutionen för mikrobiologi, Umeå universitet, Umeå (engelsk)
Opponent
Veileder
Merknad

retroaktiv registrering

Tilgjengelig fra: 2013-02-06 Laget: 2013-02-05 Sist oppdatert: 2018-03-15bibliografisk kontrollert

Open Access i DiVA

Fulltekst mangler i DiVA

Andre lenker

Forlagets fulltekstPubMed

Personposter BETA

Hamid, NiviaGustavsson, AnnaPersson, CathrineFällman, Maria

Søk i DiVA

Av forfatter/redaktør
Hamid, NiviaGustavsson, AnnaPersson, CathrineFällman, Maria
Av organisasjonen
I samme tidsskrift
Microbial Pathogenesis

Søk utenfor DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric

doi
pubmed
urn-nbn
Totalt: 243 treff
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf