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Small-molecule sensing in plants is dominated by chemical-induced dimerization modules. In the abscisic acid (ABA) system, allosteric receptors recruit phosphatase effectors and achieve nM in vivo responses from µM receptor–ligand interactions. This sensitivity amplification could enable ABA receptors to serve as generic scaffolds for designing small-molecule sensors. To test this, we screened collections of mutant ABA-receptors against 2,726 drugs and other ligands and identified 553 sensors for 6.6% of these ligands. The mutational patterns indicate strong selection for ligand-specific binding pockets. We used these data to develop a sensor design pipeline and isolated sensors for multiple plant natural products, 2,4,6-trinitrotoluene (TNT), and “forever” per- and polyfluoroalkyl substances (PFAS). Thus, the ABA sensor system enables design and isolation of small-molecule sensors with broad chemical scope and antibody-like simplicity. 

SMAX1-LIKE (SMXL) proteins in plants are cellular signaling hubs, many of which are posttranslationally regulated by karrikins from smoke, the plant hormones strigolactones (SLs), and/or cues such as light and nutrients. SMXL proteins control diverse aspects of growth, development, and environmental adaptation in plants through transcriptional corepression and interactions with transcriptional regulator proteins. In flowering plants, the SMXL family comprises four phylogenetic clades with different roles. Functions of the aSMAX1 clade include control of germination and seedling development, while the SMXL78 clade controls shoot architecture. We investigated how SMXL roles are specified inArabidopsis thaliana.Through promoter-swapping experiments, we found thatSMXL7can partially replicateSMAX1function, butSMAX1cannot replaceSMXL7. This implies that the distinct roles of these genes are primarily due to differences in protein sequences rather than expression patterns. To determine which part of SMXL proteins specifies downstream control, we tested a series of protein chimeras and domain deletions of SMAX1 and SMXL7. We found an N-terminal region that is necessary and sufficient to specify control of germination, seedling growth, or axillary branching. We screened 158 transcription factors (TFs) for interactions with SMAX1 and SMXL7 in yeast two-hybrid assays. The N-terminal domain was necessary and/or sufficient for most of the 33 potential protein–protein interactions that were identified for SMAX1. This finding unlocks different ways to engineer plant growth control through cross-wiring SMXL regulatory “input” and developmental “output” domains from different clades and lays a foundation for understanding how functional differences evolved in the SMXL family. 

Abstract SUPPRESSOR OF MAX2 1 (SMAX1) and SMAX1-LIKE (SMXL) proteins comprise a family of plant growth regulators that includes downstream targets of the karrikin (KAR)/KAI2 ligand (KL) and strigolactone (SL) signaling pathways. Following the perception of KAR/KL or SL signals by α/β hydrolases, some types of SMXL proteins are polyubiquitinated by an E3 ubiquitin ligase complex containing the F-box protein MORE AXILLARY GROWTH2 (MAX2)/DWARF3 (D3), and proteolyzed. Because SMXL proteins interact with TOPLESS (TPL) and TPL-related (TPR) transcriptional co-repressors, SMXL degradation initiates changes in gene expression. This simplified model of SMXL regulation and function in plants must now be revised in light of recent discoveries. It has become apparent that SMXL abundance is not regulated by KAR/KL or SL alone, and that some SMXL proteins are not regulated by MAX2/D3 at all. Therefore, SMXL proteins should be considered as signaling hubs that integrate multiple cues. Here we review the current knowledge of how SMXL proteins impose transcriptional regulation of plant development and environmental responses. SMXL proteins can bind DNA directly and interact with transcriptional regulators from several protein families. Multiple mechanisms of downstream genetic control by SMXL proteins have been identified recently that do not involve the recruitment of TPL/TPR, expanding the paradigm of SMXL function. 

ABSTRACT SMAX1-LIKE (SMXL) proteins are transcriptional co-repressors that regulate many aspects of plant growth and development. Proteins from the SMAX1- and SMXL78-clades of this family are targeted for degradation after karrikin or strigolactone perception, triggering downstream responses. We investigated how SMXL proteins control development.SMXL7can partially replicateSMAX1function in seeds and seedlings, butSMAX1cannot replaceSMXL7in shoot branching control. Therefore, the distinct roles of these genes arise from differences in protein activity more than expression. Analysis of chimeras and domain deletions of SMAX1 and SMXL7 proteins revealed that an N-terminal domain is necessary and sufficient to specify developmental functions. We screened 158 transcription factors for interactions with SMAX1. The N-terminal domain is necessary and/or sufficient for the majority of candidate interactions. These discoveries enable cross-wiring of karrikin and strigolactone control of plant development and lay a foundation for understanding how SMXL proteins evolved functional differences.