A Family of Bacterial Cysteine Protease Type III Effectors Utilizes Acylation-dependent and -independent Strategies to Localize to Plasma Membranes

Dowen, Robert H.; Engel, J; Shao, Feng; Ecker, Joseph R.; Dixon, Jack E.

doi:10.1074/jbc.m900519200

Cited by 95 publications

(117 citation statements)

References 51 publications

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“…4). Localization of RPS5 to the PM is consistent with previous work showing that AvrPphB becomes myristoylated inside host cells and is targeted to the PM (Dowen et al, 2009). AvrPphB catalyzes the cleavage of several related protein kinases in Arabidopsis in addition to PBS1, including BIK1 (Zhang et al, 2010).…”

Section: Pm Association Is Required For Rps5 Functionsupporting

confidence: 75%

Structure-Function Analysis of the Coiled-Coil and Leucine-Rich Repeat Domains of the RPS5 Disease Resistance Protein

2012

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The Arabidopsis (Arabidopsis thaliana) RESISTANCE TO PSEUDOMONAS SYRINGAE5 (RPS5) disease resistance protein mediates recognition of the Pseudomonas syringae effector protein AvrPphB. RPS5 belongs to the coiled-coil-nucleotide-binding site-leucine-rich repeat (CC-NBS-LRR) family and is activated by AvrPphB-mediated cleavage of the protein kinase PBS1. Here, we present a structure-function analysis of the CC and LRR domains of RPS5 using transient expression assays in Nicotiana benthamiana. We found that substituting the CC domain of RPS2 for the RPS5 CC domain did not alter RPS5 specificity and only moderately reduced its ability to activate programmed cell death, suggesting that the CC domain does not play a direct role in the recognition of PBS1 cleavage. Analysis of an RPS5-super Yellow Fluorescent Protein fusion revealed that RPS5 localizes to the plasma membrane (PM). Alanine substitutions of predicted myristoylation (glycine-2) and palmitoylation (cysteine-4) residues affected RPS5 PM localization, protein stability, and function in an additive manner, indicating that PM localization is essential to RPS5 function. The first 20 amino acids of RPS5 were sufficient for directing super Yellow Fluorescent Protein to the PM. C-terminal truncations of RPS5 revealed that the first four LRR repeats are sufficient for inhibiting RPS5 autoactivation; however, the complete LRR domain was required for the recognition of PBS1 cleavage. Substitution of the RPS2 LRR domain resulted in the autoactivation of RPS5, indicating that the LRR domain must coevolve with the NBS domain. We conclude that the RPS5 LRR domain functions to suppress RPS5 activation in the absence of PBS1 cleavage and promotes RPS5 activation in its presence.

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Section: Pm Association Is Required For Rps5 Functionsupporting

confidence: 75%

Structure-Function Analysis of the Coiled-Coil and Leucine-Rich Repeat Domains of the RPS5 Disease Resistance Protein

2012

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“…Myristoylation is a eukaryotic-specific posttranslational modification that often is used in combination with palmitoylation to target proteins to the plasma membrane (49). Multiple P. syringae T3Es acylated and localized to the plasma membrane once delivered into plant cell by the type III secretion system (50)(51)(52)(53)(54).…”

Section: Hr Triggered By Avrxv3 In Tomato Is Also Dependent On Its Pumentioning

confidence: 99%

Pseudomonas syringae type III effector HopAF1 suppresses plant immunity by targeting methionine recycling to block ethylene induction

Washington

Mukhtar

Finkel

et al. 2016

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

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HopAF1 is a type III effector protein of unknown function encoded in the genomes of several strains of Pseudomonas syringae and other plant pathogens. Structural modeling predicted that HopAF1 is closely related to deamidase proteins. Deamidation is the irreversible substitution of an amide group with a carboxylate group. Several bacterial virulence factors are deamidases that manipulate the activity of specific host protein substrates. We identified Arabidopsis methylthioadenosine nucleosidase proteins MTN1 and MTN2 as putative targets of HopAF1 deamidation. MTNs are enzymes in the Yang cycle, which is essential for the high levels of ethylene biosynthesis in Arabidopsis. We hypothesized that HopAF1 inhibits the host defense response by manipulating MTN activity and consequently ethylene levels. We determined that bacterially delivered HopAF1 inhibits ethylene biosynthesis induced by pathogenassociated molecular patterns and that Arabidopsis mtn1 mtn2 mutant plants phenocopy the effect of HopAF1. Furthermore, we identified two conserved asparagines in MTN1 and MTN2 from Arabidopsis that confer loss of function phenotypes when deamidated via site-specific mutation. These residues are potential targets of HopAF1 deamidation. HopAF1-mediated manipulation of Yang cycle MTN proteins is likely an evolutionarily conserved mechanism whereby HopAF1 orthologs from multiple plant pathogens contribute to disease in a large variety of plant hosts.

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“…Although S-palmitoylation has been detected in several hundred human proteins (48,49), the only bacterial proteins besides GobX known to receive this reversible lipidation are the type III-translocated effectors SspH2, SseI, and SifA from Salmonella (47,50) and a family of cysteine proteases from Pseudomonas syringae (51,52). There are, however, several notable differences between these effectors and GobX.…”

Section: Discussionmentioning

confidence: 98%

Host Cell-catalyzed S-Palmitoylation Mediates Golgi Targeting of the Legionella Ubiquitin Ligase GobX

Lin

Doms²,

Cheng³

et al. 2015

Journal of Biological Chemistry

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Background: During intracellular replication, Legionella pneumophila produces effector proteins that exploit host cell processes. Results:The effector GobX is an E3 ubiquitin ligase that targets to the Golgi where it is post-translationally modified with a palmitoyl moiety. Conclusion: Bacteria can hijack host-mediated S-palmitoylation for subcellular localization of their effectors. Significance: Pharmacological inhibition of S-palmitoylation enzymes may interfere with effector function and thus microbial virulence.

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A Family of Bacterial Cysteine Protease Type III Effectors Utilizes Acylation-dependent and -independent Strategies to Localize to Plasma Membranes

Cited by 95 publications

References 51 publications

Structure-Function Analysis of the Coiled-Coil and Leucine-Rich Repeat Domains of the RPS5 Disease Resistance Protein

Structure-Function Analysis of the Coiled-Coil and Leucine-Rich Repeat Domains of the RPS5 Disease Resistance Protein

Pseudomonas syringae type III effector HopAF1 suppresses plant immunity by targeting methionine recycling to block ethylene induction

Host Cell-catalyzed S-Palmitoylation Mediates Golgi Targeting of the Legionella Ubiquitin Ligase GobX

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