Luciferase Complementation Assay for Protein‐Protein Interactions in Plants

Zhou, Zhaoyang; Bi, Guozhi; Zhou, Jian‐Min

doi:10.1002/cppb.20066

Cited by 102 publications

(68 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Split‐luciferase assays were performed as described previously (Zhao & Zhou, 2020) with minor modifications. Briefly, A. tumefaciens strains EHA105 containing the desired constructs were infiltrated into fully expanded N. benthamiana leaves.…”

Section: Methodsmentioning

confidence: 99%

“…The pSPYNE-35S-cb and pSPYCE-35S-cb plasmids were described previously (Zhang et al, 2017). For split-luciferase assays, the NbNTRC and 2-Cys Prx were engineered into BamHI and SalI-digested pCAMBIA1300-CCLuc (pCCL) and pCAMBIA1300-NLuc (pNL) vectors (Zhao & Zhou, 2020), respectively, to generate the NbNTRC-cLUC and 2-Cys Prx-nLUC constructs.…”

Section: Plasmid Constructionmentioning

confidence: 99%

See 1 more Smart Citation

Barley stripe mosaic virus γb protein disrupts chloroplast antioxidant defenses to optimize viral replication

et al. 2021

View full text Add to dashboard Cite

The plant antioxidant system plays important roles in response to diverse abiotic and biotic stresses. However, the effects of virus infection on host redox homeostasis and how antioxidant defense pathway is manipulated by viruses remain poorly understood. We previously demonstrated that the Barley stripe mosaic virus (BSMV) γb protein is recruited to the chloroplast by the viral αa replicase to enhance viral replication. Here, we show that BSMV infection induces chloroplast oxidative stress. The versatile γb protein interacts directly with NADPH‐dependent thioredoxin reductase C (NTRC), a core component of chloroplast antioxidant systems. Overexpression of NbNTRC significantly impairs BSMV replication in Nicotiana benthamiana plants, whereas disruption of NbNTRC expression leads to increased viral accumulation and infection severity. To counter NTRC‐mediated defenses, BSMV employs the γb protein to competitively interfere with NbNTRC binding to 2‐Cys Prx. Altogether, this study indicates that beyond acting as a helicase enhancer, γb also subverts NTRC‐mediated chloroplast antioxidant defenses to create an oxidative microenvironment conducive to viral replication.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Plasmid Constructionmentioning

confidence: 99%

Barley stripe mosaic virus γb protein disrupts chloroplast antioxidant defenses to optimize viral replication

et al. 2021

View full text Add to dashboard Cite

show abstract

“…BIR3 and BAK1 cDNA and CSA1 and CHS3 genomic coding sequences were amplified with the primers listed in STable 2 and classically cloned into the KpnI SalI restriction sites of the vectors pCAMBIA-NLuc and pCAMBIA-CLuc for expression in Nicotiana benthamiana . The Split-Luciferase assays were performed as described in Zhou et al (2018).…”

Section: Methodsmentioning

confidence: 99%

The TIR-NBS-LRR protein CSA1 is required for autoimmune cell death in Arabidopsis pattern recognition co-receptorbak1andbir3mutants

Schulze

Ehinger

et al. 2021

Preprint

View full text Add to dashboard Cite

The BRI1-associated kinase BAK1/SERK3 is a positive regulator of multiple leucine rich receptor kinase-mediated signaling pathways including pattern triggered immunity (PTI). Absence or overexpression of BAK1 leads to spontaneous cell death formation. BAK1-interacting receptors (BIR) constitutively interact with BAK1, and plants lacking or overexpressing BIR proteins phenocopy the cell death symptoms observed in bak1 knock outs or overexpressors. In the interactome of BIR3, the TIR-NBS-LRR protein CONSTITUTIVE SHADE-AVOIDANCE 1 (CSA1) was identified by mass spectrometry. CSA1 physically interacts with BIR proteins and can be detected in complexes with BAK1. Direct interaction was shown only for CSA1 with BIR proteins but not BAK1. Double mutant bak1 bir3 genotypes develop strong dwarfism and cell death symptoms that are dependent on EDS1 and salicylic acid. Loss of CSA1 blocks bak1 and bak1 bir3-mediated cell death formation thus demonstrating that CSA1 is causal for this type of cell death. We propose that CSA1 guards BIR proteins and initiates autoimmune cell death that is observed when BAK1 BIR complexes are impaired. Our findings reveal how cell death in the absence of BAK1 and BIR3 is executed and links BAK1, a common co-receptor of many pattern recognition receptors, to NLR proteins typically implicated in effector-triggered immunity.

show abstract

“…Luciferase complementation assay. The luciferase complementation assay was conducted as described previously 97 . GS3.1 was fused to the N-terminus of luciferase (nLUC) and OsATG8b was fused with C-terminus of luciferase (cLUC).…”

Section: Methodsmentioning

confidence: 99%

A rice QTL GS3.1 regulates grain size through metabolic-flux distribution between flavonoid and lignin metabolons without affecting stress tolerance

Zhang

et al. 2021

Commun Biol

View full text Add to dashboard Cite

Grain size is a key component trait of grain weight and yield. Numbers of quantitative trait loci (QTLs) have been identified in various bioprocesses, but there is still little known about how metabolism-related QTLs influence grain size and yield. The current study report GS3.1, a QTL that regulates rice grain size via metabolic flux allocation between two branches of phenylpropanoid metabolism. GS3.1 encodes a MATE (multidrug and toxic compounds extrusion) transporter that regulates grain size by directing the transport of p-coumaric acid from the p-coumaric acid biosynthetic metabolon to the flavonoid biosynthetic metabolon. A natural allele of GS3.1 was identified from an African rice with enlarged grains, reduced flavonoid content and increased lignin content in the panicles. Notably, the natural allele of GS3.1 caused no alterations in other tissues and did not affect stress tolerance, revealing an ideal candidate for breeding efforts. This study uncovers insights into the regulation of grain size though metabolic-flux distribution. In this way, it supports a strategy of enhancing crop yield without introducing deleterious side effects on stress tolerance mechanisms.

show abstract

Luciferase Complementation Assay for Protein‐Protein Interactions in Plants

Cited by 102 publications

References 19 publications

Barley stripe mosaic virus γb protein disrupts chloroplast antioxidant defenses to optimize viral replication

Barley stripe mosaic virus γb protein disrupts chloroplast antioxidant defenses to optimize viral replication

The TIR-NBS-LRR protein CSA1 is required for autoimmune cell death in Arabidopsis pattern recognition co-receptorbak1andbir3mutants

A rice QTL GS3.1 regulates grain size through metabolic-flux distribution between flavonoid and lignin metabolons without affecting stress tolerance

Contact Info

Product

Resources

About