The C3 domain of<i>Pasteurella multocida</i>toxin is the minimal domain responsible for activation of G<sub>q</sub>‐dependent calcium and mitogenic signaling

Aminova, Leila R.; Luo, Shuhong; Bannai, Yuka; Ho, Mengfei; Wilson, Brenda A.

doi:10.1110/ps.083445408

Cited by 26 publications

(41 citation statements)

References 32 publications

(48 reference statements)

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“…Similarly, purified TcsLΔ18, although not toxic to cells after microinjection, was able to glucosylate soluble Rac in vitro (23). The toxic activity of PMT was also significantly decreased, but not completely abolished, when the C1 domain was deleted (14,26). We believe that the high amount of RID 2721-3099 -GFP produced in the cell after transfection was sufficient to locate its cellular binding partner, despite not being directly targeted to the plasma membrane.…”

Section: Discussionmentioning

confidence: 85%

Identification of a conserved membrane localization domain within numerous large bacterial protein toxins

Geissler

Tungekar²,

Satchell

2010

Proc. Natl. Acad. Sci. U.S.A.

112

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Vibrio cholerae is the causative agent of the diarrheal disease cholera. Many virulence factors contribute to intestinal colonization and disease including the Multifunctional Autoprocessing RTX toxin (MARTX Vc ). The Rho-inactivation domain (RID) of MARTX Vc is responsible for inactivating the Rho-family of small GTPases, which leads to depolymerization of the actin cytoskeleton. Based on a deletion analysis of RID to determine the minimal functional domain, we have identified a subdomain at the N terminus of RID that is homologous to the membrane targeting C1 domain of Pasteurella multocida toxin. A GFP fusion to this subdomain from RID colocalized with a plasma membrane marker when transiently expressed within HeLa cells and can be found in the membrane fraction following subcellular fractionation. This C1-like subdomain is present in multiple families of bacterial toxins, including all of the clostridial glucosyltransferase toxins and various MARTX toxins. GFP-fusions to these homologous domains are also membrane associated, indicating that this is a conserved membrane localization domain (MLD). We have identified three residues (Y23, S68, R70) as necessary for proper localization of one but not all MLDs. In addition, we found that substitution of the RID MLD with the MLDs from two different effector domains from the Vibrio vulnificus MARTX toxin restored RID activity, indicating that there is functional overlap between these MLDs. This study describes the initial recognition of a family of conserved plasma membrane-targeting domains found in multiple large bacterial toxins.bacterial toxin | multifunctional autoprocessing RTX toxin | PMT | structural modeling | Vibrio cholerae B acterial pathogens use a wide array of secreted proteins to intoxicate mammalian cells and mediate disease. Many effectors target a specific organelle, necessitating strategies to traffic to certain subcellular locations after entry into the cytosol. For example, the cytolethal distending toxin active subunit, CdtB, uses a short N-terminal nuclear localization sequence attached to traffic to the nucleus (1). Similarly, many Type III secretion effectors contain distinct targeting signals: BteA uses a 130 aa N-terminal sequence to target lipid rafts (2); ExoS and YopE use internal 21 aa domains to associate with the host membrane (3); and ExoU uses a C-terminal sequence to localize to the membrane (4). For each of these effectors, subcellular localization is determined by a specific aa sequence separate from the catalytic site.The multifunctional-autoprocessing RTX toxins (MARTX) are large bacterial toxins composed of multiple effector domains released by autoprocessing upon translocation into the host cell cytosol (5). The best characterized member of this family is the MARTX of Vibrio cholerae (MARTX Vc ), which is predicted to be 4545 aa in length (6, 7). MARTX Vc carries three effector domains delivered by autoprocessing of which two function to disrupt the actin cytoskeleton. G-actin is covalently cross-linked into oligomers by th...

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Section: Discussionmentioning

confidence: 85%

Identification of a conserved membrane localization domain within numerous large bacterial protein toxins

Geissler

Tungekar²,

Satchell

2010

Proc. Natl. Acad. Sci. U.S.A.

112

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“…Interestingly, PMT shares similarity with CNFs in the N-terminal binding and translocation domains. However, the C-terminal catalytic domain of CNFs has no obvious structural similarity with PMT, as evidenced by comparison of the crystal structure of the C-terminal C3 domain of PMT (19), harboring the minimal intracellular activity domain (20), with that of the catalytic domain of CNF1 (33) (Fig. S6 A and B).…”

Section: Discussionmentioning

confidence: 99%

“…A C-terminal fragment of PMT, which consists of 3 domains, covering amino acids 569 to 1285, has been crystallized (19). The structure revealed that the C-terminal C3 domain (residues 1105-1285), which defines the minimal intracellular activity domain responsible for activation of calcium and mitogenic signaling (20), resembles that of a papain-like protease. The domain harbors a catalytic triad characteristic of thiol proteases, consisting of the essential amino acids cysteine-1165 (17,18), histidine-1205 (21) and aspartic acid-1220.…”

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confidence: 99%

Pasteurella multocida toxin activation of heterotrimeric G proteins by deamidation

Orth

Preuß

Fester

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

103

123

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Pasteurella multocida toxin is a major virulence factor of Pasteurella multocida, which causes pasteurellosis in men and animals and atrophic rhinitis in rabbits and pigs. The Ϸ145 kDa protein toxin stimulates various signal transduction pathways by activating heterotrimeric G proteins of the G␣ q, G␣i, and G␣12/13 families by using an as yet unknown mechanism. Here, we show that Pasteurella multocida toxin deamidates glutamine-205 of G␣ i2 to glutamic acid. Therefore, the toxin inhibits the intrinsic GTPase activity of G␣ i and causes persistent activation of the G protein. A similar modification is also evident for G␣ q, but not for the closely related G␣ 11, which is not a substrate of Pasteurella multocida toxin. Our data identify the ␣-subunits of heterotrimeric G proteins as the direct molecular target of Pasteurella multocida toxin and indicate that the toxin does not act like a protease, which was suggested from its thiol protease-like catalytic triad, but instead causes constitutive activation of G proteins by deamidase activity.bacterial protein toxin ͉ Gi protein ͉ posttranslational modification ͉ transglutaminase

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“…Mutation analysis suggested that PMT may be an enzyme toxin with the Cys-His-Asp catalytic triad. More recently, the last 180 amino acids, which encompass the C3 domain, was defined as the minimum domain sufficient to stimulate both calcium-dependent and mitogenic signaling pathways [94] .…”

Section: P Multocida Toxin Pmtmentioning

confidence: 99%

Functions and Regulatory Mechanisms of Gq-Signaling Pathways

Mizuno

Itoh²

2009

Neurosignals

150

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Gq family members of heterotrimeric G protein activate β isoforms of phospholipase C that hydrolyzes phosphatidylinositol phosphate to diacylglycerol and inositol trisphosphate, leading to the protein kinase C activation and intracellular Ca²⁺ mobilization, respectively. To understand the functions and regulatory mechanisms of Gq-signaling pathways, we first introduce the Gαq-interacting proteins, which function as the effectors and the modulators of Gq. Next, we describe the Pasteurella multocida toxin and YM-254890, which are useful tools to investigate Gq signaling as activator and inhibitor, respectively. Finally, we discuss the physiological function of Gq in developmental brain, especially in neural progenitor cells.

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The C3 domain ofPasteurella multocidatoxin is the minimal domain responsible for activation of G_q‐dependent calcium and mitogenic signaling

Cited by 26 publications

References 32 publications

Identification of a conserved membrane localization domain within numerous large bacterial protein toxins

Identification of a conserved membrane localization domain within numerous large bacterial protein toxins

Pasteurella multocida toxin activation of heterotrimeric G proteins by deamidation

Functions and Regulatory Mechanisms of Gq-Signaling Pathways

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The C3 domain ofPasteurella multocidatoxin is the minimal domain responsible for activation of Gq‐dependent calcium and mitogenic signaling

Cited by 26 publications

References 32 publications

Identification of a conserved membrane localization domain within numerous large bacterial protein toxins

Identification of a conserved membrane localization domain within numerous large bacterial protein toxins

Pasteurella multocida toxin activation of heterotrimeric G proteins by deamidation

Functions and Regulatory Mechanisms of Gq-Signaling Pathways

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The C3 domain ofPasteurella multocidatoxin is the minimal domain responsible for activation of G_q‐dependent calcium and mitogenic signaling