Pathogen Protein Modularity Enables Elaborate Mimicry of a Host Phosphatase

Li, Hui; Wang, Jinlong; Kuan, Tung; Tang, Bozeng; Liu, Feng; Wang, Jiuyu; Cheng, Zhi; Sklenář, Jan; Derbyshire, Paul; Hulin, Michelle T.; Li, Yufei; Zhai, Yi; Hou, Yingnan; MacLean, Dan; Menke, Frank L. H.; Wang, Yanli; Ma, Wenbo

doi:10.2139/ssrn.4299767

Cited by 4 publications

(7 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Previous studies of another P. infestans WY/LWY effector, PSR2, demonstrated the importance of WY domains in directly binding to its host target Protein Phosphatase 2A (PP2A) (He et al 2019;Li et al 2023). Studies of mammalian PP2A and PP1c proteins have shown they share several targets in common, suggesting functional redundancy (Wlodarchak and Xing 2016;Gil and Vagnarelli 2019;Sontag et al 2008).…”

Section: Discussionmentioning

confidence: 99%

“…A subset of RxLR effectors contain one or several tandem WY domains at their C-terminus, which are important for host target specificity and effector function (Zhang et al 2019;Li et al 2023). WY domains are found in diverse oomycete species and are characterised by an 𝛼helical fold with a buried hydrophobic core (Mukhi et al 2020).…”

Section: Introductionmentioning

confidence: 99%

“…An additional class of WY effectors, called LWY effectors, carry a conserved leucine (L) residue prior to the canonical WY residues. This additional leucine is thought to be important for the structural organisation of tandem WY domains in LWY effectors, such as PSR2, creating joint-like structures that could allow the flexibility for conformational change upon interaction with other proteins, including host targets (Zhang et al 2019;Li et al 2023;He et al 2019). Intriguingly, WY domains can be decorated with additional protein-interaction motifs to aid target selectivity.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The WY domain of an RxLR effector drives interactions with a host target phosphatase to mimic host regulatory proteins and promotePhytophthora infestansinfection

Bentham¹,

Wang²,

Trusch³

et al. 2023

Preprint

View full text Add to dashboard Cite

Plant pathogens manipulate the cellular environment of the host to facilitate infection and colonization, often leading to plant diseases. To accomplish this, many specialized pathogens secrete virulence proteins called effectors into the host cell, which subvert processes such as immune signalling, gene transcription, and host metabolism. Phytophthora infestans, the causative agent of potato late blight, employs an expanded repertoire of RxLR effectors with WY domains to manipulate the host through direct interaction with protein targets. However, our understanding of the molecular mechanisms underlying the interactions between WY effectors and their host targets remains limited. In this study, we performed a structural and biophysical characterization of the P. infestans WY effector, Pi04314, in complex with the potato Protein Phosphatase 1-c (PP1c). We elucidate how Pi04314 uses a WY domain and a specialised C-terminal loop carrying a KVxF motif that interact with conserved surfaces on PP1c, known to be used by host regulatory proteins for guiding function. Through biophysical and in planta analyses, we demonstrate that Pi04314 WY or KVxF mutants lose their ability to bind PP1c. The loss of PP1c binding correlates with a reduced capacity to re-localize PP1c from the nucleolus and a decrease in lesion size in plant infection assays. This study provides insights into the manipulation of plant hosts by pathogens, revealing how effectors exploit key regulatory interfaces in host proteins to modify their function and facilitate disease.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The WY domain of an RxLR effector drives interactions with a host target phosphatase to mimic host regulatory proteins and promotePhytophthora infestansinfection

Bentham¹,

Wang²,

Trusch³

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Protein purification, immunoprecipitation, sample preparation, liquid chromatography followed by tandem mass spectrometry (LC-MS/MS), and data analysis were previously described in full detail (64). Briefly proteins identified in immunoaffinity-enriched samples were measured with data dependent method on high resolution LC-MS systems, Orbitrap Fusion (Thermo Ltd).…”

Section: Methodsmentioning

confidence: 99%

The blast effector Pwl2 is a virulence factor that modifies the cellular localisation of host protein HIPP43 to suppress immunity

Were,

Yan,

Foster

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

The rice blast fungusMagnaporthe oryzaesecretes a battery of effector proteins during host infection to suppress plant immunity and promote disease. Among these effectors, the MAX effector-Pwl2 was first identified as a host specificity determinant for infection of weeping lovegrass (Eragrostis curvula). However, its biological activity has remained unknown. Here we show that thePWL2gene is ubiquitous in all host-limited forms ofM. oryzaeand has undergone substantial copy number expansion indicating that it is a core effector of the blast fungus. We used CRISPR/Cas9-mediated gene editing to delete all three copies ofPWL2and generate apwl2null mutant inM. oryzaestrain Guy11. This resulted in gain-of-virulence towards weeping lovegrass, but a reduction in the severity of disease symptoms on rice. To further investigate the virulence mechanism of Pwl2, we showed that transgenic rice and barley lines constitutively expressingPWL2display suppression of reactive oxygen species generation and increased susceptibility to infection byM. oryzae. Also, we identify the barley and rice heavy metal-binding isoprenylated protein HIPP43 as a target of the Pwl2 effector. Transgenic lines overexpressing HIPP43 exhibit attenuated defense responses and increased susceptibility to blast infection, consistent with the hypothesis that Pwl2 binds HIPP43 to modulate immunity. This binding is coupled to changes in cellular localisation of these proteins. Taken together, our results provide evidence that Pwl2 is a virulence factor inM. oryzaethat acts by suppressing host immunity through binding and re-localising HIPP43.

show abstract

“…In addition to the RXLR motif, many Phytophthora cytoplasmic effectors also contain a conserved Glu-Glu-Arg (EER) motif, immediately downstream of the RXLR, which has again been implicated in effector delivery into host cells (Whisson et al 2007). Moreover, a ‘WY’ domain represents a conserved structural fold within the C-terminal half of many RXLR effectors that contributes to host target specificity (Boutemy et al 2011; Win et al 2012; Li et al 2023; Bentham et al 2023). Interestingly, a number of expressed effectors from Bremia lactucae that are predicted to contain the WY structural fold and are recognised by host resistance proteins only contain the EER motif (Wood et al 2020).…”

Section: Introductionmentioning

confidence: 99%

Proteolytic processing of both RXLR and EER motifs in oomycete effectors

Xu,

Wang,

Wang

et al. 2024

Preprint

View full text Add to dashboard Cite

Arg-any amino acid-Leu-Arg (RXLR) effectors are central oomycete virulence factors that target diverse host proteins and processes to suppress plant immunity. Relatively little is known about how they are processed post-translationally before delivery into host cells. Proteolytic cleavage at the RXLR motif was observed to occur prior to secretion in all Phytophthora infestans effectors tested, suggesting it is a general rule, and was observed to occur between the leucine and the second arginine. There was no cleavage of a naturally occurring second RXLR motif in a structured region of Pi21388/AvrBlb1, or one introduced at a similar position in effector Pi04314, in keeping with the motif being positionally constrained, potentially to disordered regions closely following the signal peptide. Remarkably, independent proteolytic cleavage of the Glu-Glu-Arg (EER) motif, often found immediately downstream of the RXLR, was also observed in diverse effectors, occurring immediately after the arginine. Expression of full-length effectors in host plant Nicotiana benthamiana revealed that, although secreted, they were poorly processed, suggesting that RXLR and EER cleavage does not occur in all eukaryotic cells. Our observations indicate that, whether possessing both RXLR and EER, or either motif alone, these effectors are likely proteolytically processed prior to secretion in all cases.

show abstract

Pathogen Protein Modularity Enables Elaborate Mimicry of a Host Phosphatase

Cited by 4 publications

References 43 publications

The WY domain of an RxLR effector drives interactions with a host target phosphatase to mimic host regulatory proteins and promotePhytophthora infestansinfection

The WY domain of an RxLR effector drives interactions with a host target phosphatase to mimic host regulatory proteins and promotePhytophthora infestansinfection

The blast effector Pwl2 is a virulence factor that modifies the cellular localisation of host protein HIPP43 to suppress immunity

Proteolytic processing of both RXLR and EER motifs in oomycete effectors

Contact Info

Product

Resources

About