Mechanism of Activation of Cholera Toxin by ADP-Ribosylation Factor (ARF): Both Low- and High-Affinity Interactions of ARF with Guanine Nucleotides Promote Toxin Activation

Biochemistry ◽  
1990 ◽  
Vol 29 (4) ◽  
pp. 855-861 ◽  
Author(s):  
David Bobak ◽  
Matthew Bliziotes ◽  
Masatoshi Noda ◽  
Su-Chen Tsai ◽  
Ronald Adamik ◽  
...  
Keyword(s):  
Cholera Toxin ◽  
Guanine Nucleotides ◽  
Adp Ribosylation ◽  
High Affinity ◽  
Adp Ribosylation Factor

scholarly journals Characterization of the human gene encoding ADP-ribosylation factor 1, a guanine nucleotide-binding activator of cholera toxin.

1992 ◽  
Vol 267 (13) ◽  
pp. 9028-9034
Author(s):  
C.M. Lee ◽  
R.S. Haun ◽  
S.C. Tsai ◽  
J Moss ◽  
M Vaughan
Keyword(s):  
Cholera Toxin ◽  
Human Gene ◽  
Nucleotide Binding ◽  
Guanine Nucleotide ◽  
Gene Encoding ◽  
Adp Ribosylation ◽  
Guanine Nucleotide Binding ◽  
Adp Ribosylation Factor

Separation of the 24 kDa substrate for botulinum C3 ADP-ribosyltransferase and the cholera toxin ADP-ribosylation factor

1988 ◽  
Vol 152 (3) ◽  
pp. 957-961 ◽  
Author(s):  
Su-Chen Tsai ◽  
Ronald Adamik ◽  
Joel Moss ◽  
Klaus Aktories
Keyword(s):  
Cholera Toxin ◽  
Adp Ribosylation ◽  
Adp Ribosyltransferase ◽  
Adp Ribosylation Factor

Effects of temperature on ADP-ribosylation factor stimulation of cholera toxin activity

Biochemistry ◽  
1993 ◽  
Vol 32 (2) ◽  
pp. 561-566 ◽  
Author(s):  
Toshihiko Murayama ◽  
Su Chen Tsai ◽  
Ronald Adamik ◽  
Joel Moss ◽  
Martha Vaughan
Keyword(s):  
Cholera Toxin ◽  
Adp Ribosylation ◽  
Effects Of Temperature ◽  
Adp Ribosylation Factor ◽  
Stimulation Of

Effects of guanine nucleotides on cholera toxin catalyzed ADP-ribosylation in rat adipocyte plasma membranes

Biochemistry ◽  
1983 ◽  
Vol 22 (26) ◽  
pp. 6291-6296 ◽  
Author(s):  
C. Bruce Graves ◽  
Nancy B. Klaven ◽  
Jay M. McDonald
Keyword(s):  
Cholera Toxin ◽  
Plasma Membranes ◽  
Guanine Nucleotides ◽  
Adp Ribosylation ◽  
Adipocyte Plasma Membranes

scholarly journals The Cholera Toxin A13 Subdomain Is Essential for Interaction with ADP-Ribosylation Factor 6 and Full Toxic Activity but Is Not Required for Translocation from the Endoplasmic Reticulum to the Cytosol

2006 ◽  
Vol 74 (4) ◽  
pp. 2259-2267 ◽  
Author(s):  
Ken Teter ◽  
Michael G. Jobling ◽  
Danielle Sentz ◽  
Randall K. Holmes
Keyword(s):  
Endoplasmic Reticulum ◽  
Cholera Toxin ◽  
Toxic Activity ◽  
Hydrophobic Domain ◽  
Vesicular Traffic ◽  
Adp Ribosylation ◽  
Quality Control Mechanism ◽  
Adp Ribosylation Factor

ABSTRACT Cholera toxin (CT) moves from the plasma membrane to the endoplasmic reticulum (ER) by retrograde vesicular traffic. In the ER, the catalytic CTA1 polypeptide dissociates from the rest of the toxin and enters the cytosol by a process that involves the quality control mechanism of ER-associated degradation (ERAD). The cytosolic CTA1 then ADP ribosylates Gsα, resulting in adenylate cyclase activation and intoxication of the target cell. It is hypothesized that the C-terminal A13 subdomain of CTA1 plays two crucial roles in the intoxication process: (i) it contains a hydrophobic domain that triggers the ERAD mechanism and (ii) it facilitates interaction with the cytosolic ADP-ribosylation factors (ARFs) that serve as allosteric activators of CTA1. In this study, we examined the role(s) of the CTA13 subdomain in CT intoxication. Full-length CTA1 constructs and truncated CTA1 constructs lacking the A13 subdomain were generated and used to conduct two-hybrid studies of interactions with ARF6, in vitro enzyme assays, in vivo toxicity assays, and in vivo processing/degradation assays. Direct, plasmid-mediated expression of CTA1 constructs in the ER or cytosol of transfected CHO cells was used to perform the in vivo assays. With these methods, we found that the A13 subdomain of CTA1 is important both for interaction with ARF6 and for full expression of enzyme activity in vivo. Surprisingly, however, the A13 subdomain was not required for ERAD-mediated passage of CTA1 from the ER to the cytosol. A possible alternative trigger for CTA1 to activate the ERAD mechanism is discussed.

scholarly journals Differential cholera-toxin- and pertussis-toxin-catalysed ADP-ribosylation of G-proteins coupled to formyl-peptide and leukotriene B4 receptors

1993 ◽  
Vol 289 (2) ◽  
pp. 469-473 ◽  
Author(s):  
T M Schepers ◽  
K R McLeish
Keyword(s):  
Cholera Toxin ◽  
G Proteins ◽  
Pertussis Toxin ◽  
Leukotriene B4 ◽  
Guanine Nucleotides ◽  
Dependent Manner ◽  
Functional Responses ◽  
Concentration Dependent Manner ◽  
Adp Ribosylation ◽  
Formyl Peptide

N-Formylmethionyl-leucyl-phenylalanine (fMet-Leu-Phe) and leukotriene B4 (LTB4) induce disparate second-messenger generation and functional responses in neutrophils and HL-60 granulocytes. Receptors for these chemoattractants couple to a common pool of G-proteins which are substrates for both pertussis-toxin- and cholera-toxin-catalysed ADP-ribosylation. The hypothesis that formyl-peptide and LTB4 receptors induce different receptor-specific conformations of activated G-proteins was tested. The ability of pertussis toxin and cholera toxin to ADP-ribosylate G(i) proteins coupled to formyl-peptide or LTB4 receptors in membranes isolated from HL-60 granulocytes was used to assess the conformational state of the alpha subunits. Cholera-toxin-catalysed ADP-ribosylation of alpha 40 (40 kDa alpha subunit) was inhibited by guanosine 5′-[beta gamma-imido]triphosphate and GDP in a concentration-dependent manner. Addition of fMet-Leu-Phe, but not LTB4, re-established cholera-toxin labelling of alpha 40 in the presence of either guanine nucleotide. In the absence of guanine nucleotides, fMet-Leu-Phe and C5a enhanced cholera-toxin-catalysed labelling of alpha 40, whereas LTB4 and platelet-activating factor had no effect. Preincubation with fMet-Leu-Phe, but not LTB4, inhibited pertussis-toxin labelling of alpha 40 in the presence of guanosine 5′-[gamma-thio]triphosphate and in the absence of guanine nucleotides. Preincubation with fMet-Leu-Phe or LTB4 enhanced pertussis-toxin labelling of alpha 40 in the presence of GDP. These data suggest that activated G(i) proteins coupled to formyl-peptide and LTB4 receptors exist in different conformations determined by the receptor with which they interact.

scholarly journals ADP-Ribosylation Factor 1 Transiently Activates High-Affinity Adaptor Protein Complex AP-1 Binding Sites On Golgi Membranes

1998 ◽  
Vol 9 (6) ◽  
pp. 1323-1337 ◽  
Author(s):  
Yunxiang Zhu ◽  
Linton M. Traub ◽  
Stuart Kornfeld
Keyword(s):  
Protein Complex ◽  
Binding Sites ◽  
Adaptor Protein ◽  
Limiting Factor ◽  
Golgi Membrane ◽  
Gtp Hydrolysis ◽  
Adp Ribosylation ◽  
High Affinity ◽  
Golgi Membranes ◽  
Adp Ribosylation Factor

Association of the Golgi-specific adaptor protein complex 1 (AP-1) with the membrane is a prerequisite for clathrin coat assembly on the trans-Golgi network (TGN). The AP-1 adaptor is efficiently recruited from cytosol onto the TGN by myristoylated ADP-ribosylation factor 1 (ARF1) in the presence of the poorly hydrolyzable GTP analog guanosine 5′-O-(3-thiotriphosphate) (GTPγS). Substituting GTP for GTPγS, however, results in only poor AP-1 binding. Here we show that both AP-1 and clathrin can be recruited efficiently onto the TGN in the presence of GTP when cytosol is supplemented with ARF1. Optimal recruitment occurs at 4 μM ARF1 and with 1 mM GTP. The AP-1 recruited by ARF1·GTP is released from the Golgi membrane by treatment with 1 M Tris-HCl (pH 7) or upon reincubation at 37°C, whereas AP-1 recruited with GTPγS or by a constitutively active point mutant, ARF1(Q71L), remains membrane bound after either treatment. An incubation performed with added ARF1, GTP, and AlFn, used to block ARF GTPase-activating protein activity, results in membrane-associated AP-1, which is largely insensitive to Tris extraction. Thus, ARF1·GTP hydrolysis results in lower-affinity binding of AP-1 to the TGN. Using two-stage assays in which ARF1·GTP first primes the Golgi membrane at 37°C, followed by AP-1 binding on ice, we find that the high-affinity nucleating sites generated in the priming stage are rapidly lost. In addition, the AP-1 bound to primed Golgi membranes during a second-stage incubation on ice is fully sensitive to Tris extraction, indicating that the priming stage has passed the ARF1·GTP hydrolysis point. Thus, hydrolysis of ARF1·GTP at the priming sites can occur even before AP-1 binding. Our finding that purified clathrin-coated vesicles contain little ARF1 supports the concept that ARF1 functions in the coat assembly process rather than during the vesicle-uncoating step. We conclude that ARF1 is a limiting factor in the GTP-stimulated recruitment of AP-1 in vitro and that it appears to function in a stoichiometric manner to generate high-affinity AP-1 binding sites that have a relatively short half-life.

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