Purification and characterization of an ADP-ribosyltransferase produced by Clostridium limosum.

Just, Ingo; Mohr, Christiane; Schallehn, G; Ménard, Luc; Didsbury, John; Vandekerckhove, Joël; Damme, Josef Van; Aktories, Klaus

doi:10.1016/s0021-9258(19)50014-x

Cited by 163 publications

(43 citation statements)

References 37 publications

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“…Because loss of rho-1 causes numerous pleiotropic developmental phenotypes and embryonic lethality (Jantsch-Plunger et al 2000), we employed the widely-used Rho inhibitor C. botulinum C3T, which ADP-ribosylates Rho (Aktories et al 2004). C3T has strong substrate specificity for Rho and has only weak activity toward other Rho family members like Rac and Cdc42 (Just et al 1992). Though effects of C3T on targets other than Rho cannot be absolutely excluded, a study in C. elegans found that C3T, rho-1 RNAi, and expression of a dominant-negative form of Rho all caused similar defects in P cell migration (Spencer et al 2001).…”

Section: Inhibition Of Rho Suppresses Activated G Qmentioning

confidence: 99%

The NCA-1 and NCA-2 Ion Channels Function Downstream of Gq and Rho To Regulate Locomotion in Caenorhabditis elegans

et al. 2017

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The heterotrimeric G protein G positively regulates neuronal activity and synaptic transmission. Previously, the Rho guanine nucleotide exchange factor Trio was identified as a direct effector of G that acts in parallel to the canonical G effector phospholipase C. Here, we examine how Trio and Rho act to stimulate neuronal activity downstream of G in the nematode Through two forward genetic screens, we identify the cation channels NCA-1 and NCA-2, orthologs of mammalian NALCN, as downstream targets of the G-Rho pathway. By performing genetic epistasis analysis using dominant activating mutations and recessive loss-of-function mutations in the members of this pathway, we show that NCA-1 and NCA-2 act downstream of G in a linear pathway. Through cell-specific rescue experiments, we show that function of these channels in head acetylcholine neurons is sufficient for normal locomotion in Our results suggest that NCA-1 and NCA-2 are physiologically relevant targets of neuronal G-Rho signaling in .

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Section: Inhibition Of Rho Suppresses Activated G Qmentioning

confidence: 99%

The NCA-1 and NCA-2 Ion Channels Function Downstream of Gq and Rho To Regulate Locomotion in Caenorhabditis elegans

et al. 2017

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“…Thus, a second injection of C3 transferase into cells previously injected with L61Rac and C3 caused the dissolution of focal complexes (data not shown). As C3 can also ribosylate and inactivate Rac in vitro [22], we could not exclude the possibility that in addition to further inhibition of Rho activity, increased C3 concentrations also suppress Rac activity in vivo. Injection of a Rac mutant lacking the C3-ribosylation site resolved this question.…”

Section: Discussionmentioning

confidence: 99%

“…To answer this question we used much higher concentrations of C3, and this led to the loss of focal complexes. Rac is also a potential substrate of C3, however, albeit a poor one [22], and the possibility had to be considered that high C3 concentrations also lead to the inactivation of Rac by ribosylation. To circumvent this problem, we constructed a mutant of constitutively active L61Rac (I39L61Rac) in which the Asn39 ribosylation site was exchanged for isoleucine.…”

Section: Rac-mediated Focal-complex Formation and Membrane Ruffling Is Independent Of Rho And Rho-kinasementioning

confidence: 99%

Interplay between Rac and Rho in the control of substrate contact dynamics

1999

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Rac functions to signal the creation of new substrate contacts at the cell front, which are associated with the induction of ruffling lamellipodia, whereas Rho serves in the maturation of existing contacts, with both contact types requiring contractility for their formation. The transition from a focal complex to a focal contact is associated with a switch to Rho-kinase dependence. Rac and Rho also influence the development of focal contacts and focal complexes, respectively, through mutually antagonistic pathways.

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“…The aim of the present study was to further characterize the role of GTPases of the Rho family in the pertussis toxin-sensitive LPA-mediated induction of Cox-2 in mesangial cells. Toxin B from Clostridium difficile [19] and C3 transferase from C. limosum [20] were used to inhibit the three members of the Rho family, Rho, Rac and Cdc42, and to interfere specifically with RhoA signalling respectively. In particular, we employed the C3-fusion toxin C2IN-C3 (where C2IN is part of the C2I toxin of C. botulinum), which is able to enter target cells readily [21,22].…”

Section: Introductionmentioning

confidence: 99%

Role of Rac and Cdc42 in lysophosphatidic acid-mediated cyclo-oxygenase-2 gene expression

et al. 2002

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The role of Rho proteins in lysophosphatidic acid (LPA)-mediated induction of cyclo-oxygenase-2 (Cox-2) was investigated in renal mesangial cells. Previous studies had shown that toxin B, an inhibitor of Rho, Rac and Cdc42, suppressed Cox-2 induction. A role for RhoA in pertussis toxin-sensitive LPA signalling was excluded with C3 transferase from Clostridium limosum, used as the fusion toxin C2IN-C3 (where C2IN is part of the C2I toxin of C. botulinum). Incubation of the cells with C2IN-C3 disrupted cytosolic actin stress fibres, but had no effect on Cox-2 induction. Similarly, activation of p42/44 mitogen-activated protein kinase (MAP kinase), an upstream step in Cox-2 induction, was inhibited by toxin B, but not affected by C2IN-C3. Upon treatment with toxin B, focal adhesion kinase and paxillin were dephosphorylated at tyrosine residues and the actin cytoskeleton was completely destroyed. An intact cytoskeleton, however, was not required for p42/44 MAP-kinase activation or Cox-2 induction, as shown by the actin-depolymerizing agent cytochalasin D. Toxin B did not influence functionality of LPA receptors, because G(i)-mediated Ca(2+) release from intracellular stores remained unchanged. Within 1 h, toxin B inactivated and translocated RhoA and Cdc42 to the cellular membranes. Within the same time frame, monoglucosylated Rac1 was degraded. Direct stimulation of Rho proteins by cytotoxic necrotizing factor type 1 (CNF1) induced Cox-2 expression, which was sensitive to inhibition of the MAP-kinase pathway by PD98059, but not to an inhibitor of RhoA kinase. By exclusion of RhoA and non-specific cytoskeletal effects, the results in the present study indicate an important role for Rac and/or Cdc42 in pertussis toxin-sensitive LPA-mediated Cox-2 induction.

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Purification and characterization of an ADP-ribosyltransferase produced by Clostridium limosum.

Cited by 163 publications

References 37 publications

The NCA-1 and NCA-2 Ion Channels Function Downstream of Gq and Rho To Regulate Locomotion in Caenorhabditis elegans

The NCA-1 and NCA-2 Ion Channels Function Downstream of Gq and Rho To Regulate Locomotion in Caenorhabditis elegans

Interplay between Rac and Rho in the control of substrate contact dynamics

Role of Rac and Cdc42 in lysophosphatidic acid-mediated cyclo-oxygenase-2 gene expression

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