Regulatory cascade involving transcriptional and N-end rule pathways in rice under submergence

Lin, Chih-Cheng; Chao, Ya-Ting; Chen, Wan-Chieh; Ho, Hao-Chung; Chou, Mei-Yi; Li, Yaru; Wu, Yulin; Yang, Hung-An; Hsieh, Hsiang; Lin, Chun-Shin; Wu, Fan; Chou, Shu‐Jen; Jen, Hao-Chung; Huang, Yuanyuan; Irene, Deli; Wu, Wen‐Jin; Wu, Jianli; Gibbs, Daniel J.; Ho, Meng-Chiao; Shih, Ming-Che

doi:10.1073/pnas.1818507116

Cited by 85 publications

(87 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…One prominent example for a protein that evades the PRT6 N‐degron pathway, despite containing the N‐terminal motif, is SUB1‐A1, which is a major determinant of submergence tolerance in rice (Fukao et al ., ; Gibbs et al ., ). For this protein, it was recently demonstrated that the C‐terminus protects it from degradation (Lin et al ., ).…”

Section: Discussionmentioning

confidence: 97%

AtERF#111/ABR1 is a transcriptional activator involved in the wounding response

et al. 2019

View full text Add to dashboard Cite

AtERF#111/ABR1 belongs to the group X of the ERF/AP2 transcription factor family (GXERFs) and is shoot specifically induced under submergence and hypoxia. It was described to be an ABA-response repressor, but our data reveal a completely different function. Surprisingly, AtERF#111 expression is strongly responsive to wounding stress. Expression profiling of ERF#111-overexpressing (OE) plants, which show morphological phenotypes like increased root hair length and number, strengthens the hypothesis of AtERF#111 being involved in the wounding response, thereby acting as a transcriptional activator of gene expression. Consistent with a potential function outside of oxygen signalling, we could not assign AtERF#111 as a target of the PRT6 N-degron pathway, even though it starts with a highly conserved N-terminal MetÀCys (MC) motif. However, the protein is unstable as it is degraded in an ubiquitin-dependent manner. Finally, direct target genes of AtERF#111 were identified by microarray analyses and subsequently confirmed by protoplast transactivation assays. The special roles of diverse members of the plant-specific GXERFs in coordinating stress signalling and wound repair mechanisms have been recently hypothesized, and our data suggest that AtERF#111 is indeed involved in these processes.For hypoxia treatments, 7-day-old seedlings grown on MS medium were used. 2 h after the onset of the photoperiod, open Petri dishes were placed into a desiccator for the indicated time periods

show abstract

Section: Discussionmentioning

confidence: 97%

AtERF#111/ABR1 is a transcriptional activator involved in the wounding response

et al. 2019

View full text Add to dashboard Cite

show abstract

“…The ERFVII proteins are highly conserved in flowering plants (Nakano et al ), and rice ERFVIIs include SUBMERGENCE(SUB)1A and SNORKEL (SKL)1 and SKL2, that have been shown to enhance tolerance to rapid or deep‐water flooding, respectively (Xu et al ; Hattori et al ). Interestingly although SUB1A protein contains Cys2 it was shown that it is not a substrate for the PRT6 N‐degron pathway in vitro (Gibbs et al ), because the C‐terminus protects the N‐terminus by directly interacting (Lin et al ), providing a possible explanation for the improved flood‐tolerance of accessions containing this locus. Two other rice ERFVIIs, 66 and 67 were shown to act downstream of SUB1A, and to be targets of N‐degron pathway‐mediated degradation (Lin et al ).…”

Section: Physiological Roles Of N‐degron Pathways In Plant Growth Andmentioning

confidence: 99%

The plant N‐degron pathways of ubiquitin‐mediated proteolysis

Holdsworth

Vicente

Sharma

et al. 2019

JIPB

View full text Add to dashboard Cite

The amino-terminal residue of a protein (or amino-terminus of a peptide following protease cleavage) can be an important determinant of its stability, through the Ubiquitin Proteasome System associated N-degron pathways. Plants contain a unique combination of N-degron pathways (previously called the N-end rule pathways) E3 ligases, PROTEOLYSIS (PRT)6 and PRT1, recognizing non-overlapping sets of amino-terminal residues, and others remain to be identified. Although only very few substrates of PRT1 or PRT6 have been identified, substrates of the oxygen and nitric oxide sensing branch of the PRT6 N-degron pathway include key nuclear-located transcription factors (ETHYLENE RESPONSE FACTOR VIIs and LITTLE ZIPPER 2) and the histone-modifying Polycomb Repressive Complex 2 component VERNALIZATION 2. In response to reduced oxygen or nitric oxide levels (and other mechanisms that reduce pathway activity) these stabilized substrates regulate diverse aspects of growth and development, including response to flooding, salinity, vernalization (cold-induced flowering) and shoot apical meristem function. The N-degron pathways show great promise for use in the improvement of crop performance and for biotechnological applications. Upstream proteases, components of the different pathways and associated substrates still remain to be identified and characterized to fully appreciate how regulation of protein stability through the amino-terminal residue impacts plant biology.

show abstract

“…In particular, some of these ERF transcription factors in rice, namely ERF66, ERF67 and SUB1A-1, all have the attributes of an N-degron, as well as a conserved N-terminal region. Despite these common features, SUB1A-1 is not targeted for degradation by the N-degron pathway, while ERF66/67 are degraded via this pathway (Lin et al, 2019). The use of truncated proteins, as well as interaction assays, indicated that the C-terminal domain of SUB1A-1 could fold back onto the N-terminal region, thus precluding recognition of the N-terminal destabilizing residue by N-degron components (Lin et al, 2019).…”

Section: Determinants Of N-degrons In Plantsmentioning

confidence: 99%

“…Despite these common features, SUB1A-1 is not targeted for degradation by the N-degron pathway, while ERF66/67 are degraded via this pathway (Lin et al, 2019). The use of truncated proteins, as well as interaction assays, indicated that the C-terminal domain of SUB1A-1 could fold back onto the N-terminal region, thus precluding recognition of the N-terminal destabilizing residue by N-degron components (Lin et al, 2019). Interestingly, the Nterminal region of SUB1A-1 is also largely unstructured (Lin et al, 2019), similarly to the RIN4 fragments, and presumably also the NOI fragments.…”

Section: Determinants Of N-degrons In Plantsmentioning

confidence: 99%

“…The use of truncated proteins, as well as interaction assays, indicated that the C-terminal domain of SUB1A-1 could fold back onto the N-terminal region, thus precluding recognition of the N-terminal destabilizing residue by N-degron components (Lin et al, 2019). Interestingly, the Nterminal region of SUB1A-1 is also largely unstructured (Lin et al, 2019), similarly to the RIN4 fragments, and presumably also the NOI fragments. It is hence possible that differences in the conformation of the different fragments released after AvrRpt2 cleavage could explain the different degradation mechanisms.…”

Section: Determinants Of N-degrons In Plantsmentioning

confidence: 99%

See 1 more Smart Citation

Differential N-end rule degradation of RIN4/NOI fragments generated by the AvrRpt2 effector protease

Goslin

Eschen‐Lippold

Naumann

et al. 2019

Preprint

View full text Add to dashboard Cite

One sentence summary: Analysis of RIN4/NOI fragments released after cleavage by the bacterial effector protease AvrRpt2 reveals a novel role of the N-end rule in the degradation of NOI-domain proteins, but not of RIN4.Author contributions: KG, AK and EG constructed plasmids used in tobacco and Arabidopsis protoplast experiments, including tFT plasmids; KG carried out tobacco transient expression experiments, immunoblots and confocal imaging, as well as all immunoblots for RIN4; LE-L conducted all NOI experiments in Arabidopsis protoplasts, including immunoblots; CN and MK constructed plasmids for in vitro arginylation assays and performed these assays; EL conducted tFT experiments; MS and RdM carried out inoculations with Pst AvrRpt2 and confocal analysis; EG, ND, JL and MW supervised the experiments; EG wrote the article with contributions of all the authors; EG agrees to serve as the author responsible for contact and ensures communication. AbstractThe protein RPM1-INTERACTING PROTEIN4 (RIN4) is a central regulator of both layers of plant immunity systems, the so-called pattern-triggered immunity (PTI) and effectortriggered immunity (ETI). RIN4 is targeted by several effectors, including the Pseudomonas syringae protease effector AvrRpt2. Cleavage of RIN4 by AvrRpt2 generates unstable RIN4 fragments, whose degradation leads to the activation of the resistance protein RPS2 (RESISTANT TO P. SYRINGAE2). Hence, identifying the determinants of RIN4 degradation is key to understanding RPS2-mediated ETI, as well as virulence functions of AvrRpt2. In addition to RIN4, AvrRpt2 cleaves host proteins from the nitrate-induced (NOI) domain family. Although cleavage of NOI-domain proteins by AvrRpt2 may contribute to PTI regulation, the (in)stability of these proteolytic fragments and the determinants that regulate their stability have not been examined. Notably, a common feature of RIN4 and of many NOI-domain protein fragments generated by AvrRpt2 cleavage is the exposure of a new N-terminal residue that is destabilizing according to the N-end rule. Using antibodies raised against endogenous RIN4, we show that the destabilization of AvrRpt2-cleaved RIN4 fragments is independent of the N-end rule pathway (recently renamed N-degron pathway). By contrast, several NOI-domain protein fragments are bona fide substrates of the N-degron pathway. The discovery of this novel set of substrates considerably expands the number of proteins targeted for degradation by this ubiquitin-dependent pathway, for which very few physiological substrates are known in plants. Our results also open new avenues of research to understand the role of AvrRpt2 in promoting bacterial virulence.

show abstract

Regulatory cascade involving transcriptional and N-end rule pathways in rice under submergence

Cited by 85 publications

References 59 publications

AtERF#111/ABR1 is a transcriptional activator involved in the wounding response

AtERF#111/ABR1 is a transcriptional activator involved in the wounding response

The plant N‐degron pathways of ubiquitin‐mediated proteolysis

Differential N-end rule degradation of RIN4/NOI fragments generated by the AvrRpt2 effector protease

Contact Info

Product

Resources

About