Search for Partner Proteins of A. thaliana Immunophilins Involved in the Control of Plant Immunity

Abstract

The involvement of plant immunophilins in multiple essential processes such as development, various ways of adapting to biotic and abiotic stresses, and photosynthesis has already been established. Previously, research has demonstrated the involvement of three immunophilin genes (AtCYP19-1/ROC3, AtFKBP65/ROF2, and AtCYP57) in the control of plant response to invasion by various pathogens. Current research attempts to identify host target proteins for each of the selected immunophilins. As a result, candidate interactors have been determined and confirmed using a yeast 2-hybrid (Y2H) system for protein–protein interaction assays. The generation of mutant isoforms of ROC3 and AtCYP57 harboring substituted amino acids in the in silico-predicted active sites became essential to achieving significant binding to its target partners. This data shows that ROF2 targets calcium-dependent lipid-binding domain-containing protein (At1g70790; AT1) and putative protein phosphatase (At2g30020; АТ2), whereas ROC3 interacts with GTP-binding protein (At1g30580; ENGD-1) and RmlC-like cupin (At5g39120). The immunophilin AtCYP57 binds to putative pyruvate decarboxylase-1 (Pdc1) and clathrin adaptor complex-related protein (At5g05010). Identified interactors confirm our previous findings that immunophilins ROC3, ROF2, and AtCYP57 are directly involved with stress response control. Further, these findings extend our understanding of the molecular functional pathways of these immunophilins.

The Arabidopsis Proteins AtNHR2A and AtNHR2B Are Multi-Functional Proteins Integrating Plant Immunity With Other Biological Processes

“…With respect to the interaction of AtNHR2A and AtNHR2B with proteins known to be important in regulation of plant immunity, we found that AtNHR2A interacted with AtPRL1, a protein that interacts with the plant defense transcription factor AtCDC5 (Palma et al, 2007), with the MAP kinase AtMPK11 (Carrasco et al, 2014), and with AtENGD-1 (Cheung et al, 2010). AtENGD-1 is a GTP-binding protein that participates in plant immunity regulating the production of ROS and interacting with the immunophilin AtCYP19, which is also important for plant defense responses against P. syringae (Cheung et al, 2010;Pogorelko et al, 2014;Abdeeva et al, 2018). We successfully validated the interaction between AtNHR2A and AtENGD-1 by semi-in vivo co-immunoprecipitation wherein His-AtENGD-1 purified from bacteria was successfully co-immunoprecipitated by AtNHR2A-GFP expressed and purified from N. benthamiana.…”

“…To further confirm the accuracy of the IP-MS method, and to gain insight into the function of AtNHR2A and AtNHR2B in plant immunity, we chose to validate the interaction between AtNHR2A or AtNHR2B with proteins known to function in plant innate immunity such as AtENGD-1, AtRPN1A, and AtCCoAMT1 (Yao et al, 2012;Abdeeva et al, 2018;Xie et al, 2018). AtENGD-1 was fused to the 6× histidine (6× His) tag and expressed and purified from E. coli.…”

Interactome of Arabidopsis Thaliana

“…According to TAIR, the genome of Arabidopsis thaliana contains 27,416 protein-coding genes ( Table S3 ). The main portion of PPIs was identified in several databases, while an additional 920 PPIs were identified through a PubMed search for studies of interactions in Arabidopsis published over the last twenty years [ 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , …”

Cyclophilin OsCYP20‐2 with a novel variant integrates defense and cell elongation for chilling response in rice

“…The major functions of cyclophilin are as foldase, scaffold and chaperon, and in deed CYP imparts on its target protein stability and activity, which participates in plant growth, abiotic and biotic stress response (Zhu, ; Fu et al , ; Pogorelko et al , ; Abdeeva et al , ). To further reveal the relationships between OsCYP20‐2 and OsFSD2, we performed a Y3H assay and showed that OsCYP20‐2 promoted OsFSD2 forming homodimers (Fig.…”