Identification of In Planta Protein–Protein Interactions Using IP-MS

Jamge, Suraj; Angenent, Gerco C.; Bemer, Marian

doi:10.1007/978-1-4939-7318-7_18

Cited by 9 publications

(6 citation statements)

References 7 publications

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“…Recently, affinity enrichment protocols have been updated and further optimized for nuclear, cytoplasmic and membrane-associated protein complexes ( Wendrich et al, 2017 ; Jamge et al, 2018 ).…”

Section: Dealing With False-positive and False-negative Resultsmentioning

confidence: 99%

Recent Trends in Plant Protein Complex Analysis in a Developmental Context

et al. 2018

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Because virtually all proteins interact with other proteins, studying protein–protein interactions (PPIs) is fundamental in understanding protein function. This is especially true when studying specific developmental processes, in which proteins often make developmental stage- or tissue specific interactions. However, studying these specific PPIs in planta can be challenging. One of the most widely adopted methods to study PPIs in planta is affinity purification coupled to mass spectrometry (AP/MS). Recent developments in the field of mass spectrometry have boosted applications of AP/MS in a developmental context. This review covers two main advancements in the field of affinity purification to study plant developmental processes: increasing the developmental resolution of the harvested tissues and moving from affinity purification to affinity enrichment. Furthermore, we discuss some new affinity purification approaches that have recently emerged and could have a profound impact on the future of protein interactome analysis in plants.

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Section: Dealing With False-positive and False-negative Resultsmentioning

confidence: 99%

Recent Trends in Plant Protein Complex Analysis in a Developmental Context

et al. 2018

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“…Besides, new studies should focus on searching for other proteins/transcriptional factors that might interact with bHLHs (via heterodimers) and regulate their functions. Approaches such as ChIP-Seq and co-immunoprecipitation, tested in different anther development stages, for instance, would clarify which genes are directly regulated by the aforementioned bHLHs and which proteins make heterodimers with them, respectively ( Jamge et al, 2018 ; Nakato and Sakata, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

bHLH-regulated routes in anther development in rice and Arabidopsis

Ortolan,

Trenz,

Delaix

et al. 2023

Genet. Mol. Biol.

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Anther development is a complex process essential for plant reproduction and crop yields. In recent years, significant progress has been made in the identification and characterization of the bHLH transcription factor family involved in anther regulation in rice and Arabidopsis, two extensively studied model plants. Research on bHLH transcription factors has unveiled their crucial function in controlling tapetum development, pollen wall formation, and other anther-specific processes. By exploring deeper into regulatory mechanisms governing anther development and bHLH transcription factors, we can gain important insights into plant reproduction, thereby accelerating crop yield improvement and the development of new plant breeding strategies. This review provides an overview of the current knowledge on anther development in rice and Arabidopsis, emphasizing the critical roles played by bHLH transcription factors in this process. Recent advances in gene expression analysis and functional studies are highlighted, as they have significantly enhanced our understanding of the regulatory networks involved in anther development.

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“…Immunoprecipitation and mass spectrometry analysis (IP‐MS) and co‐immunoprecipitation (Co‐IP) assays were performed as detailed before (Jamge et al ., 2018 ; Jin et al ., 2020 ). Briefly, total proteins, extracted from the derived plant tissues, were centrifuged for 15 min (18 200 g) at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

“…The immunoprecipitated protein complexes were eluted by boiling the beads for 5 min in 2× SDS‐PAGE sample buffer, which were then separated using 12% SDS‐PAGE. After silver staining (Jamge et al ., 2018 ), the desired protein bands were excised and subjected to LC–MS/MS analysis. For Co‐IP assays, the immunoprecipitated proteins separated by SDS‐PAGE were transferred to nitrocellulose membranes, followed by immunoblotting with appropriate antibodies (Table S4 ) (Jin et al ., 2020 ).…”

Section: Methodsmentioning

confidence: 99%

Appropriate reduction of importin‐α gene expression enhances yellow dwarf disease resistance in common wheat

Wang,

Zhang,

Wang

et al. 2023

Plant Biotechnology Journal

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SummaryBarley yellow dwarf viruses (BYDVs) cause widespread damage to global cereal crops. Here we report a novel strategy for elevating resistance to BYDV infection. The 17K protein, a potent virulence factor conserved in BYDVs, interacted with barley IMP‐α1 and ‐α2 proteins that are nuclear transport receptors. Consistently, a nuclear localization signal was predicted in 17K, which was found essential for 17K to be transported into the nucleus and to interact with IMP‐α1 and ‐α2. Reducing HvIMP‐α1 and ‐α2 expression by gene silencing attenuated BYDV‐elicited dwarfism, accompanied by a lowered nuclear accumulation of 17K. Among the eight common wheat CRISPR mutants with two to four TaIMP‐α1 and ‐α2 genes mutated, the triple mutant α1aaBBDD/α2AAbbdd and the tetra‐mutant α1aabbdd/α2AAbbDD displayed strong BYDV resistance without negative effects on plant growth under field conditions. The BYDV resistance exhibited by α1aaBBDD/α2AAbbdd and α1aabbdd/α2AAbbDD was correlated with decreased nuclear accumulation of 17K and lowered viral proliferation in infected plants. Our work uncovers the function of host IMP‐α proteins in BYDV pathogenesis and generates the germplasm valuable for breeding BYDV‐resistant wheat. Appropriate reduction of IMP‐α gene expression may be broadly useful for enhancing antiviral resistance in agricultural crops and other economically important organisms.

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Identification of In Planta Protein–Protein Interactions Using IP-MS

Cited by 9 publications

References 7 publications

Recent Trends in Plant Protein Complex Analysis in a Developmental Context

Recent Trends in Plant Protein Complex Analysis in a Developmental Context

bHLH-regulated routes in anther development in rice and Arabidopsis

Appropriate reduction of importin‐α gene expression enhances yellow dwarf disease resistance in common wheat

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