Biology subjects

Mooney, B. C.

Publications and source records attributed to Mooney, B. C..

3 recordsLinked to original sources

bioRxiv2024

Bioengineering secreted proteases converts divergent Rcr3 orthologs and paralogs into extracellular immune co-receptors

Secreted immune proteases Rcr3 and Pip1 of tomato are both inhibited by Avr2 from the fungal plant pathogen Cladosporium fulvum but only Rcr3 act as a decoy co-receptor that detects Avr2 in the presence of the Cf-2 immune receptor. Here, we identified crucial residues from tomato Rcr3 required for Cf-2-mediated signalling and bioengineered various proteases to trigger Avr2/Cf-2 dependent immunity. Despite substantial divergences in Rcr3 orthologs from eggplant and tobacco, only minimal alterations were sufficient to trigger Avr2/Cf-2-triggered immune signalling. Tomato Pip1, by contrast, was bioengineered with 16 Rcr3-specific residues to initiate Avr2/Cf-2-triggered immune signalling. These residues cluster on one side next to the substrate binding groove, indicating a potential Cf-2 interaction site. Our findings also revealed that Rcr3 and Pip1 have distinct substrate preferences determined by two variant residues and that both are suboptimal for binding Avr2. This study advances our understanding of Avr2 perception and opens avenues to bioengineer proteases to broaden pathogen recognition in other crops.

plant biology↗

bioRxiv2023

Repression of pattern-triggered immune responses by hypoxia

Biotic and abiotic stresses frequently co-occur in nature, yet, relatively little is known about how plants co-ordinate the response to combined stresses. Previous research has shown that protein degradation by the ubiquitin/proteasome system is central to the regulation of multiple independent stress response pathways in plants. The Arg/N-degron pathway is a subset of the ubiquitin/proteasome system that targets proteins based on their N-termini and has been specifically implicated in the responses to biotic and abiotic stresses, including hypoxia via accumulation of ERF-VII transcription factors, which orchestrate the onset of the hypoxia response program. Here, we investigated the role of the Arg/N-degron pathway in mediating the crosstalk between coinciding abiotic and biotic stresses using hypoxia treatments and the flg22 elicitor of pattern-triggered immunity (PTI), respectively. We uncovered a link between the transcriptional responses of plants to hypoxia and flg22. Combined hypoxia/flg22 treatments showed that hypoxia represses the flg22 transcriptional program, as well as the expression of pattern recognition receptors, MAPK signalling and callose deposition during PTI, through mechanisms that are mostly independent from the ERF-VIIs. These findings aid understanding of the trade-offs between plant responses to combined abiotic/biotic stresses in the context of our efforts to increase crop resilience to global climate change. Our results also show that the well-known repressive effect of hypoxia on innate immunity in animals also applies to plants. Significance statementUnderstanding how plants regulate the crosstalk between stress response pathways is key to our efforts to increase crop resilience and mitigate yield losses caused by global climate change. Despite the urgency to do so, relatively little is known about how plants respond to combined stresses, which frequently occur in nature. Here, we show that the hypoxia response program and the basal layer of plant immunity (pattern-triggered immunity or PTI) share components. Our data also show that hypoxia represses several key aspects of PTI, a situation akin to that discovered in animals decades ago. These findings have implications for our ability to develop resilient crops by limiting the negative trade-offs that exist between hypoxia response and immunity.

plant biology↗

bioRxiv2023

BIG participates in the Arg/N-degron pathways and the hypoxia response in Arabidopsis thaliana.

BIG (also known as DOC1 and TIR3) is an 0.5 MDa protein that has been associated with multiple important functions in signalling and development through forward genetic screens in Arabidopsis thaliana. However, the biochemical function(s) of BIG are unknown. Here, we investigated whether BIG plays a role in the Arg/N-degron pathways, protein regulatory mechanisms in which substrate protein fate is influenced by the N-terminal (Nt) residue. In Arabidopsis, PROTEOLYSIS1 (PRT1) is an E3 ligase with specificity for aromatic amino acids, whereas PROTEOLYSIS6 (PRT6) targets basic N-terminal residues. We crossed a big loss-of-function allele to prt6 and prt1 mutants and examined the stability of protein substrates. Stability of model N-degron pathway substrates was enhanced in prt6-1 big-2 and prt1-1 big-2 relative to the respective single mutants. Abundance of the PRT6 physiological substrates, HYPOXIA RESPONSIVE ERF (HRE)2 and VERNALIZATION (VRN)2 was similarly increased in prt6 big double mutants, without increase in transcripts. Accordingly, hypoxia marker expression was enhanced in prt6 big double mutants, in a manner requiring arginyltransferase activity and RAP-type ERFVII transcription factors. Transcriptomic analysis of roots not only demonstrated synergistically increased expression of a plethora of hypoxia responsive genes in the double mutant relative to prt6 but also revealed other roles for PRT6 and BIG, including regulation of suberin deposition through both ERFVII-dependent and independent mechanisms, respectively. Our results show that BIG acts together with PRT6 to regulate the hypoxia response and wider processes. Significance StatementThe N-degron pathways are a group of protein regulatory mechanisms that play important roles in plant growth, development, and response to biotic and abiotic stresses. Despite rapid progress in the last decade, key enzymatic components of the pathways remain to be identified. BIG (also known as DOC1 and TIR3) is a protein of approximately 0.5 MDa, associated with multiple, distinct roles in plants but the precise biochemical functions of this protein have remained enigmatic until now. Here we identify BIG as a new component of plant N-degron pathways that acts together with the N-recognin E3 ligase PROTEOLYSIS6 (PRT6) to control the hypoxia response and other functions in Arabidopsis thaliana.

plant biology↗