Identification of a Molecular Target for the Yersinia Protein Kinase A

Navarro, Lorena; Koller, Antonius; Nordfelth, Roland; Wolf‐Watz, Hans; Taylor, Susan S.; Dixon, Jack E.

doi:10.1016/j.molcel.2007.04.025

Cited by 89 publications

(93 citation statements)

References 44 publications

(80 reference statements)

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“…In contrast to VopA that modifies the nucleotide binding site with a neutral acetyl group, YpkA phosphorylates a serine residue found in the proximity of the GTP binding site (16). Phosphorylation of Ser-47 on G␣ q is predicted to reduce its affinity for GTP, thereby inactivating the G-protein.…”

Section: Discussionmentioning

confidence: 99%

VopA Inhibits ATP Binding by Acetylating the Catalytic Loop of MAPK Kinases

Trosky

Mukherjee

et al. 2007

Journal of Biological Chemistry

108

107

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The bacterial pathogen Vibrio parahemeolyticus manipulates host signaling pathways during infections by injecting type III effectors. One of these effectors, Vibrio outer protein A (VopA), inhibits MAPK signaling via a novel mechanism, distinct from those described for other bacterial toxins, that disrupts this signaling pathway. VopA is an acetyltransferase that potently inhibits MAPK signaling pathways not only by preventing the activation of MAPK kinases (MKKs) but also by inhibiting the activity of activated MKKs. VopA acetylates a conserved lysine found in the catalytic loop of all kinases and blocks the binding of ATP, but not ADP, on the MKKs, resulting in an inactive phosphorylated kinase. Acetylation of this conserved lysine inhibits kinase activity by a new mechanism of regulation that has not been observed previously. Identifying the target of VopA reveals a way that the reversible post-translational modification of lysine acetylation can be used to regulate the activity of an enzyme.Vibrio parahemeolyticus is a marine bacterium and causative agent of gastroenteritis associated with the consumption of contaminated seafood. It is endemic to Southeast Asia and is the leading cause of gastroenteritis in Japan (1). Sequencing of the V. parahemeolyticus genome revealed the presence of two type III secretion systems each encoded in separate pathogenicity islands (2). Type III secretion systems are used by bacterial pathogens to inject effector proteins into the cytoplasm of the host cells (3). Upon delivery into the host cytosol, the effectors manipulate host signaling to gain an advantage for the infecting pathogen. The bacterial effectors are proposed to usurp or mimic eukaryotic activity that is then used to target and deregulate normal functions in the host cell by either up-or downregulating signaling pathways (4). However, the bacterial effector proteins must initially be kept in a quiescent state within the bacterium. This is accomplished by a variety of methods, including their association with a chaperone in the bacterial cytosol, their specificity for a substrate, and/or their requirement for an activator or modification that is only provided by a eukaryotic source (4, 5).VopA (Vibrio outer protein A) is an effector found within one of the V. parahemeolyticus pathogenicity islands, and is a member of a family of type III effectors called YopJ-like proteins that are found in a wide variety of pathogens including other animal pathogens, plant pathogens, and the plant symbiont Rhizobium (5-7). The founding member of the family, YopJ from Yersinia spp., inhibits the mitogen-activated protein kinase (MAPK) 3 and NFB signaling cascades within the host cell, thereby inhibiting the host's innate immune response (7). All YopJ-like proteins contain a catalytic triad, and it was recently shown that YopJ functions as an acetyltransferase (8 -10). It has been speculated that other members of the family of YopJ-like proteins might also possess this activity (8). YopJ was demonstrated to inhibit the MAPK and NFB...

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Section: Discussionmentioning

confidence: 99%

VopA Inhibits ATP Binding by Acetylating the Catalytic Loop of MAPK Kinases

Trosky

Mukherjee

et al. 2007

Journal of Biological Chemistry

108

107

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“…YopE acts as a GAP for RhoA and Rac and induces their inactivation (von Pawel-Rammingen et al 2000;Andor et al 2001). YopO/YpkA is a multidomain protein with a GDI-like binding domain (Prehna et al 2006) and an actin-binding domain (Dukuzumuremyi et al 2000), and an N-terminal serine/threonine kinase domain, which inhibits Gaq signaling pathways by direct phosphorylation of Gaq (Navarro et al 2007). YopT is a member of cysteine proteases with invariant Cys/His/Asp residues (Shao et al 2002) that causes rounding of host cells by disruption of the actin cytoskeleton .…”

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confidence: 99%

The polybasic region of Rho GTPases defines the cleavage by Yersinia enterocolitica outer protein T (YopT)

Fueller

Schmidt

2008

Protein Science

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Pathogenic Yersinia strains evade the innate immune responses of the host by producing effector proteins (Yersinia outer proteins [Yops]), which are directly injected into mammalian cells by a type III secretion system (TTSS). One of these effector proteins (YopT) disrupts the actin cytoskeleton of the host cell resulting in cell rounding. YopT is a cysteine protease that cleaves Rho proteins directly upstream of the post-translationally modified cysteine. Thereby, it releases the GTPases from the membrane leading to inactivation. Small GTPases are modified by isoprenylation of the cysteine of the CAAX box, cleavage of the -AAX tripeptide, and methylation of the cysteine. We have shown that isoprenylation and the endoproteolytic cleavage of the tripeptide of Rho GTPases are essential for YopT-induced cleavage, whereas carboxyl methylation is not required. In the present study, we posttranslationally modified RhoA, Rac, Cdc42, and several mutants in vitro and characterized the YopTinduced cleavage with recombinant YopT. We show that farnesylated RhoA is a preferred substrate of YopT compared with the geranylgeranylated GTPase. Geranylgeranylated RhoA, however, is the preferred substrate for YopT-catalyzed cleavage with a threefold faster turnover rate over Rac and Cdc42. Moreover, our data indicate that the composition of the polybasic region of the GTPases defines the specificity and efficiency of the YopT-induced cleavage, and that a space between the polybasic stretch of amino acids at the C terminus and the CAAX box enhances the turnover rate of YopT-catalyzed cleavage.

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“…YpkA of Yersinia inhibits GTP binding through phosphorylation of the host heterotrimeric G protein G␣ q (49). Similarly, but with less mechanistic detail, a Salmonella SPI2 effector called SteC is known to manipulate actin using its kinase activity (8).…”

Section: Fig 2 Host Cell-specific Modulation Of Cellular Processes mentioning

confidence: 99%

Global Impact of Salmonella Pathogenicity Island 2-secreted Effectors on the Host Phosphoproteome

Imami

Bhavsar

et al. 2013

Molecular & Cellular Proteomics

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During the late stages of infection, Salmonella secretes numerous effectors through a type III secretion system that is encoded within Salmonella pathogenicity island 2 (SPI2). Despite the importance of SPI2 as a major virulence factor leading to the systemic spread of the bacteria and diseases, a global view of its effects on host responses is still lacking. Here, we measured global impacts of SPI2 effectors on the host phosphorylation and protein expression levels in RAW264.7 and in HeLa cells, as macrophage and nonphagocytic models of infection. We observe that SPI2 effectors differentially modulate the host phosphoproteome and cellular processes (e.g. protein trafficking, cytoskeletal regulation, and immune signaling) in a host cell-dependent manner. Our unbiased approach reveals the involvement of many previously unrecognized proteins, including E3 ligases (HERC4, RanBP2, and RAD18), kinases (CDK, SIK3, and WNK1), and histones (H2B1F, H4, and H15), in late stages of Salmonella infection. Furthermore, from this phosphoproteome analysis and other quantitative screens, we identified HSP27 as a direct in vitro and in vivo molecular target of the only type III secreted kinase, SteC. Using biochemical and cell biological assays, we demonstrate that SteC phosphorylates multiple sites in HSP27 and induces actin rearrangement through this protein. Together, these results provide a broader landscape of host players contributing to specific processes/pathways mediated by SPI2 effectors than was previously appreciated. Molecular &

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Identification of a Molecular Target for the Yersinia Protein Kinase A

Cited by 89 publications

References 44 publications

VopA Inhibits ATP Binding by Acetylating the Catalytic Loop of MAPK Kinases

VopA Inhibits ATP Binding by Acetylating the Catalytic Loop of MAPK Kinases

The polybasic region of Rho GTPases defines the cleavage by Yersinia enterocolitica outer protein T (YopT)

Global Impact of Salmonella Pathogenicity Island 2-secreted Effectors on the Host Phosphoproteome

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