cAMP imaging of cells treated with pertussis toxin, cholera toxin, and anthrax edema toxin.

The enzymatic activity of the three most studied bacterial toxins that increase the cytosolic cAMP level: pertussis toxin (PT), cholera toxin (CT), and anthrax edema toxin (ET), was imaged by fluorescence videomicroscopy. Three different cell lines were transfected with a fluorescence resonance ener...

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Main Authors: Dal Molin, F, Zornetta, I, Puhar, A, Tonello, F, Zaccolo, M, Montecucco, C
פורמט: Journal article
שפה:English
יצא לאור: 2008
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author Dal Molin, F
Zornetta, I
Puhar, A
Tonello, F
Zaccolo, M
Montecucco, C
author_facet Dal Molin, F
Zornetta, I
Puhar, A
Tonello, F
Zaccolo, M
Montecucco, C
author_sort Dal Molin, F
collection OXFORD
description The enzymatic activity of the three most studied bacterial toxins that increase the cytosolic cAMP level: pertussis toxin (PT), cholera toxin (CT), and anthrax edema toxin (ET), was imaged by fluorescence videomicroscopy. Three different cell lines were transfected with a fluorescence resonance energy transfer biosensor based on the PKA regulatory and catalytic subunits fused to CFP and YFP, respectively. Real-time imaging of cells expressing this cAMP biosensor provided time and space resolved pictures of the toxins action. The time course of the PT-induced cAMP increase suggests that its active subunit enters the cytosol more rapidly than that deduced by biochemical experiments. ET generated cAMP concentration gradients decreasing from the nucleus to the cell periphery. On the contrary, CT, which acts on the plasma membrane adenylate cyclase, did not. The potential of imaging methods in studying the mode of entry and the intracellular action of bacterial toxins is discussed.
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spelling oxford-uuid:823e0b0c-948d-465d-9b86-bc69c2ef27162022-03-26T21:36:03ZcAMP imaging of cells treated with pertussis toxin, cholera toxin, and anthrax edema toxin.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:823e0b0c-948d-465d-9b86-bc69c2ef2716EnglishSymplectic Elements at Oxford2008Dal Molin, FZornetta, IPuhar, ATonello, FZaccolo, MMontecucco, CThe enzymatic activity of the three most studied bacterial toxins that increase the cytosolic cAMP level: pertussis toxin (PT), cholera toxin (CT), and anthrax edema toxin (ET), was imaged by fluorescence videomicroscopy. Three different cell lines were transfected with a fluorescence resonance energy transfer biosensor based on the PKA regulatory and catalytic subunits fused to CFP and YFP, respectively. Real-time imaging of cells expressing this cAMP biosensor provided time and space resolved pictures of the toxins action. The time course of the PT-induced cAMP increase suggests that its active subunit enters the cytosol more rapidly than that deduced by biochemical experiments. ET generated cAMP concentration gradients decreasing from the nucleus to the cell periphery. On the contrary, CT, which acts on the plasma membrane adenylate cyclase, did not. The potential of imaging methods in studying the mode of entry and the intracellular action of bacterial toxins is discussed.
spellingShingle Dal Molin, F
Zornetta, I
Puhar, A
Tonello, F
Zaccolo, M
Montecucco, C
cAMP imaging of cells treated with pertussis toxin, cholera toxin, and anthrax edema toxin.
title cAMP imaging of cells treated with pertussis toxin, cholera toxin, and anthrax edema toxin.
title_full cAMP imaging of cells treated with pertussis toxin, cholera toxin, and anthrax edema toxin.
title_fullStr cAMP imaging of cells treated with pertussis toxin, cholera toxin, and anthrax edema toxin.
title_full_unstemmed cAMP imaging of cells treated with pertussis toxin, cholera toxin, and anthrax edema toxin.
title_short cAMP imaging of cells treated with pertussis toxin, cholera toxin, and anthrax edema toxin.
title_sort camp imaging of cells treated with pertussis toxin cholera toxin and anthrax edema toxin
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AT zornettai campimagingofcellstreatedwithpertussistoxincholeratoxinandanthraxedematoxin
AT puhara campimagingofcellstreatedwithpertussistoxincholeratoxinandanthraxedematoxin
AT tonellof campimagingofcellstreatedwithpertussistoxincholeratoxinandanthraxedematoxin
AT zaccolom campimagingofcellstreatedwithpertussistoxincholeratoxinandanthraxedematoxin
AT montecuccoc campimagingofcellstreatedwithpertussistoxincholeratoxinandanthraxedematoxin