General Information:

Id: 3,006
Diseases: Pseudomonas aeruginosa diseases
pathogen-host system
Pseudomonas aeruginosa/mammalia
reporter system based on Bla/CCF2 enzyme/substrate combination
article
Reference: Verove J et al.(2012) Injection of Pseudomonas aeruginosa Exo Toxins into Host Cells Can Be Modulated by Host Factors at the Level of Translocon Assembly and/or Activity. PLoS ONE 7 [PMID: 22299042]

Interaction Information:

Comment To analyse T3SS translocon function in vivo a simple translocation reporter system based on the activity of beta--lactamase that cleaves a fluorescent substrate was used. The resulting chimeric proteins ExoS-Bla and ExoY-Bla were efficiently translocated into several cell lines such as the epithelial cell line A549, the B cell line BJAB, the T cell line Jurkat, and the differentiated promyelocytic HL-60 and promonocytic U937. No injection of the reporter fusion could be observed with undifferentiated HL-60 or U937 cells.
Formal Description
Interaction-ID: 27370

process

T3SS

increases_transport of

gene/protein

exoS

in A459, BJAB, Jurkat, differentiated HL-60 and U937 cells
Comment To analyse T3SS translocon function in vivo a simple translocation reporter system based on the activity of beta--lactamase that cleaves a fluorescent substrate was used. The resulting chimeric proteins ExoS-Bla and ExoY-Bla were efficiently translocated into several cell lines such as the epithelial cell line A549, the B cell line BJAB, the T cell line Jurkat, and the differentiated promyelocytic HL-60 and promonocytic U937. No injection of the reporter fusion could be observed with undifferentiated HL-60 or U937 cells.
Formal Description
Interaction-ID: 27371

process

T3SS

increases_transport of

gene/protein

exoY

in A459, BJAB, Jurkat, differentiated HL-60 and U937 cells
Comment To analyse T3SS translocon function in vivo a simple translocation reporter system based on the activity of beta--lactamase that cleaves a fluorescent substrate was used. The resulting chimeric proteins ExoS-Bla and ExoY-Bla were efficiently translocated into several cell lines such as the epithelial cell line A549, the B cell line BJAB, the T cell line Jurkat, and the differentiated promyelocytic HL-60 and promonocytic U937. No injection of the reporter fusion could be observed with undifferentiated HL-60 or U937 cells.
Formal Description
Interaction-ID: 27372

process

T3SS

NOT affects transport of

gene/protein

exoS

in undifferentiated HL-60 or U937 cells
Comment To analyse T3SS translocon function in vivo a simple translocation reporter system based on the activity of beta--lactamase that cleaves a fluorescent substrate was used. The resulting chimeric proteins ExoS-Bla and ExoY-Bla were efficiently translocated into several cell lines such as the epithelial cell line A549, the B cell line BJAB, the T cell line Jurkat, and the differentiated promyelocytic HL-60 and promonocytic U937. No injection of the reporter fusion could be observed with undifferentiated HL-60 or U937 cells.
Formal Description
Interaction-ID: 27373

process

T3SS

NOT affects transport of

gene/protein

exoY

in undifferentiated HL-60 or U937 cells
Comment Translocon insertion into HL-60 membranes is independent of cell differentiation. Even though the injection of effectors occurs only in differentiated cells, the association of PopB/D translocators to membranes occurs equally well in both non-permissive and permissive cells. This suggests that the pore assembly and/or functionality may be regulated by host cell factors.
Formal Description
Interaction-ID: 27374

complex/PPI

T3SS translocon complex

is localized in

cellular component

host cell plasma membrane

in undifferentiated, or VD3-differentiated HL-60 cells
Comment Cholesterol-rich membrane microdomains have been implicated in invasion of P. aeruginosa. Depletion of cholesterol from lipid rafts by methyl-beta-cyclodextrin abolished the translocation of the reporter ExoS-Bla in a dose-dependent manner. Co-localisation of PopB with the lipid raft marker flotillin is further supporting the role of lipid rafts in the translocation process.
Formal Description
Interaction-ID: 27377

drug/chemical compound

Methyl-beta-cyclodextrin

decreases transport of

gene/protein

exoS

by cholesterol removal of cultured cells
Comment The actin network appears to play a major role in the translocation process in the VD3-differentiated-HL-60 model as evidenced by the action of cytochalasin D and latrunculin B, two actin-depolymerizing agents which were found to markedly inhibit the injection of ExoS-Bla.
Formal Description
Interaction-ID: 27378

affects transport of

gene/protein

exoS

Comment The actin network appears to play a major role in the translocation process in the VD3-differentiated-HL-60 model as evidenced by the action of cytochalasin D and latrunculin B, two actin-depolymerizing agents which were found to markedly inhibit the injection of ExoS-Bla.
Formal Description
Interaction-ID: 27379

drug/chemical compound

Cytochalasin D

decreases transport of

gene/protein

exoS

into VD3-dHL-60 cells
Comment The actin network appears to play a major role in the translocation process in the VD3-differentiated-HL-60 model as evidenced by the action of cytochalasin D and latrunculin B, two actin-depolymerizing agents which were found to markedly inhibit the injection of ExoS-Bla.
Formal Description
Interaction-ID: 27380

drug/chemical compound

Cytochalasin D

decreases_activity of

Comment The actin network appears to play a major role in the translocation process in the VD3-differentiated-HL-60 model as evidenced by the action of cytochalasin D and latrunculin B, two actin-depolymerizing agents which were found to markedly inhibit the injection of ExoS-Bla.
Formal Description
Interaction-ID: 27405

drug/chemical compound

Latrunculin B

decreases transport of

gene/protein

exoS

into VD3-dHL-60 cells
Drugbank entries Show/Hide entries for Latrunculin B
Comment The actin network appears to play a major role in the translocation process in the VD3-differentiated-HL-60 model as evidenced by the action of cytochalasin D and latrunculin B, two actin-depolymerizing agents which were found to markedly inhibit the injection of ExoS-Bla.
Formal Description
Interaction-ID: 27406

drug/chemical compound

Latrunculin B

decreases_activity of

Drugbank entries Show/Hide entries for Latrunculin B
Comment Two specific PI3-kinase inhibitors, wortmannin and LY-294002 inhibited the translocation of ExoS-Bla, suggesting the involvement of PI3K signalling pathway in the translocon activity.
Formal Description
Interaction-ID: 27420

affects transport of

gene/protein

exoS

Comment Two specific PI3-kinase inhibitors, wortmannin and LY-294002 inhibited the translocation of ExoS-Bla, suggesting the involvement of PI3K signalling pathway in the translocon activity.
Formal Description
Interaction-ID: 27423

drug/chemical compound

Wortmannin

decreases transport of

gene/protein

exoS

into VD3-dHL-60 cells
Comment Two specific PI3-kinase inhibitors, wortmannin and LY-294002 inhibited the translocation of ExoS-Bla, suggesting the involvement of PI3K signalling pathway in the translocon activity.
Formal Description
Interaction-ID: 27424

drug/chemical compound

Wortmannin

decreases_activity of

Comment Two specific PI3-kinase inhibitors, wortmannin and LY-294002 inhibited the translocation of ExoS-Bla, suggesting the involvement of PI3K signalling pathway in the translocon activity.
Formal Description
Interaction-ID: 27425

drug/chemical compound

LY294002

decreases transport of

gene/protein

exoS

into VD3-dHL-60 cells
Comment Two specific PI3-kinase inhibitors, wortmannin and LY-294002 inhibited the translocation of ExoS-Bla, suggesting the involvement of PI3K signalling pathway in the translocon activity.
Formal Description
Interaction-ID: 27426

drug/chemical compound

LY294002

decreases_activity of

Comment ExoS-Bla injection into VD3-dHL-60 was abolished by genistein, a large spectrum tyrosine kinase inhibitor, by PP2, an inhibitor of the Src-kinase(s), and by PF-5732208, an inhibitor of focal adhesion tyrosine kinases, FAK and Pyk2.
Formal Description
Interaction-ID: 27427

drug/chemical compound

Genistein

decreases transport of

gene/protein

exoS

into VD3-dHL-60 cells
Drugbank entries Show/Hide entries for Genistein
Comment ExoS-Bla injection into VD3-dHL-60 was abolished by genistein, a large spectrum tyrosine kinase inhibitor, by PP2, an inhibitor of the Src-kinase(s), and by PF-5732208, an inhibitor of focal adhesion tyrosine kinases, FAK and Pyk2.
Formal Description
Interaction-ID: 27428

drug/chemical compound

PP2 (kinase inhibitor)

decreases transport of

gene/protein

exoS

into VD3-dHL-60 cells
Comment ExoS-Bla injection into VD3-dHL-60 was abolished by genistein, a large spectrum tyrosine kinase inhibitor, by PP2, an inhibitor of the Src-kinase(s), and by PF-5732208, an inhibitor of focal adhesion tyrosine kinases, FAK and Pyk2.
Formal Description
Interaction-ID: 27429

drug/chemical compound

PF-5732208

decreases transport of

gene/protein

exoS

into VD3-dHL-60 cells
Comment ExoS-Bla injection into VD3-dHL-60 was abolished by genistein, a large spectrum tyrosine kinase inhibitor, by PP2, an inhibitor of the Src-kinase(s), and by PF-5732208, an inhibitor of focal adhesion tyrosine kinases, FAK and Pyk2.
Formal Description
Interaction-ID: 27430

drug/chemical compound

PF-5732208

decreases_activity of

gene/protein

PTK2

into VD3-dHL-60 cells
Drugbank entries Show/Hide entries for PTK2
Comment ExoS-Bla injection into VD3-dHL-60 was abolished by genistein, a large spectrum tyrosine kinase inhibitor, by PP2, an inhibitor of the Src-kinase(s), and by PF-5732208, an inhibitor of focal adhesion tyrosine kinases, FAK and Pyk2.
Formal Description
Interaction-ID: 27431

drug/chemical compound

PF-5732208

decreases_activity of

gene/protein

PTK2B

Drugbank entries Show/Hide entries for PTK2B
Comment The association of PopB translocator with membranes of VD3-dHL-60 cells was not impeded when cells were treated with the different inhibitors. Thus, the insertion of the translocon proteins can be uncoupled from the translocation process per se.
Formal Description
Interaction-ID: 27432

drug/chemical compound

Cytochalasin D

NOT decreases_folding of

complex/PPI

T3SS translocon complex

at the host plasma membrane (VD3-dHL-60 cells)
Comment The association of PopB translocator with membranes of VD3-dHL-60 cells was not impeded when cells were treated with the different inhibitors. Thus, the insertion of the translocon proteins can be uncoupled from the translocation process per se.
Formal Description
Interaction-ID: 27437

drug/chemical compound

Latrunculin B

NOT decreases_folding of

complex/PPI

T3SS translocon complex

at the host plasma membrane (VD3-dHL-60 cells)
Drugbank entries Show/Hide entries for Latrunculin B
Comment The association of PopB translocator with membranes of VD3-dHL-60 cells was not impeded when cells were treated with the different inhibitors. Thus, the insertion of the translocon proteins can be uncoupled from the translocation process per se.
Formal Description
Interaction-ID: 27438

drug/chemical compound

Wortmannin

NOT decreases_folding of

complex/PPI

T3SS translocon complex

at the host plasma membrane (VD3-dHL-60 cells)
Comment The association of PopB translocator with membranes of VD3-dHL-60 cells was not impeded when cells were treated with the different inhibitors. Thus, the insertion of the translocon proteins can be uncoupled from the translocation process per se.
Formal Description
Interaction-ID: 27439

drug/chemical compound

LY294002

NOT decreases_folding of

complex/PPI

T3SS translocon complex

at the host plasma membrane (VD3-dHL-60 cells)
Comment The association of PopB translocator with membranes of VD3-dHL-60 cells was not impeded when cells were treated with the different inhibitors. Thus, the insertion of the translocon proteins can be uncoupled from the translocation process per se.
Formal Description
Interaction-ID: 27440

drug/chemical compound

Genistein

NOT decreases_folding of

complex/PPI

T3SS translocon complex

at the host plasma membrane (VD3-dHL-60 cells)
Drugbank entries Show/Hide entries for Genistein
Comment The association of PopB translocator with membranes of VD3-dHL-60 cells was not impeded when cells were treated with the different inhibitors. Thus, the insertion of the translocon proteins can be uncoupled from the translocation process per se.
Formal Description
Interaction-ID: 27441

drug/chemical compound

PP2 (kinase inhibitor)

NOT decreases_folding of

complex/PPI

T3SS translocon complex

at the host plasma membrane (VD3-dHL-60 cells)
Comment The association of PopB translocator with membranes of VD3-dHL-60 cells was not impeded when cells were treated with the different inhibitors. Thus, the insertion of the translocon proteins can be uncoupled from the translocation process per se.
Formal Description
Interaction-ID: 27442

drug/chemical compound

PF-5732208

NOT decreases_folding of

complex/PPI

T3SS translocon complex

at the host plasma membrane (VD3-dHL-60 cells)
Comment Cholesterol-rich membrane microdomains have been implicated in invasion of P. aeruginosa. Depletion of cholesterol from lipid rafts by methyl-beta-cyclodextrin abolished the translocation of the reporter ExoS-Bla in a dose-dependent manner. Co-localisation of PopB with the lipid raft marker flotillin is further supporting the role of lipid rafts in the translocation process.
Formal Description
Interaction-ID: 27554

drug/chemical compound

Methyl-beta-cyclodextrin

decreases_quantity of

cellular component

membrane raft