ABI5 Expression Research Articles

The BHLH Transcriptional Factor PIF4 Competes with the R2R3-MYB Transcriptional Factor MYB75 to Fine-Tune Seeds Germination under High Glucose Stress

It is known that the high level of sugar including glucose suppresses seed germination through ABA signal. ABI5 is an essential component to mediate ABA-dependent seed germination inhibition, but underlying mechanism needs more investigation. Previous study demonstrated the PIF4 activated the expression of ABI5 to suppress seed germination in darkness. Here we reported that PIF4 also mediated the seed germination inhibition through ABI5 under high concentration of glucose treatment. Furthermore, we found that PIF4 interacted with PAP1, the central factor to control anthocyanin biosynthesis. Such interaction was confirmed in vitro and in planta. Biochemical and physiological analysis revealed that PAP1 bond the promoter of ABI5 to suppress its expression, thus enhanced seed germination under high concentration of glucose treatment. Specially, PAP1 competed with PIF4 to antagonize the activation of PIF4 on ABI5 expression, thus promoted seed germination under high glucose treatment. Given these, we uncover a novel role for PIF4 and PAP1 in controlling seed germination under high glucose treatment, and reveal their antagonistic mechanism by which coordinates ABI5 expression to control seed germination in response to the glucose signal.

Arabidopsis F-box protein At1g08710 interacts with transcriptional protein ADA2b and imparts drought stress tolerance by negatively regulating seedling growth

Plants experience abiotic stresses throughout their life cycle and accordingly respond to tide over the unfavorable conditions. Plants adopt to drought stress through various molecular, biochemical, physiological and cellular processes. F-box protein subunit of the Skp1-Cullin-F-box (SCF) E3 ubiquitin ligases plays crucial role in imparting specificity for selective degradation of target proteins. Here we report the function of Arabidopsis F-box protein At1g08710 in drought stress adaptation. F-box protein is a constituent of SCF complex as it is shown interacting with ASK1 and Cullin 1. F-box protein localizes in both nucleus and membrane. F-box gene transcript is highly accumulated in root and altered in response to drought stress conditions. F-box protein interacts with a transcriptional co-activator protein ADA2b. F-box mutant plants growth is better under drought stress conditions compared to the wild type. Accumulation of H2O2 and malondialdehyde (MDA) content is reduced in mutant plants. Drought responsive genes RD29A, RD22, ABI3 expression is induced in F-box mutant plants. These results indicate F-box protein At1g08710 role in drought stress adaptation in Arabidopsis thaliana.

Expression of storage lipid biosynthesis transcription factors and enzymes in Jatropha curcas L. cell suspension cultures and seeds

The oleaginous Jatropha curcas has been proposed as a promising source for biodiesel production in seed or potentially by in vitro production in cell cultures. However, little is known concerning the optimal growth conditions and the transcription of key factors and enzymes involved in the biosynthesis of storage lipids in J. curcas cell cultures. Additional knowledge is also needed for these factors in seeds. Here, we assess target gene expression in endosperm cells in planta and endosperm-derived cell suspension cultures (EDCCs). Endosperm cells were taken from three representative seed developmental stages, and cell suspensions were grown from these samples. Glucose, nitrogen, and abscisic acid concentrations were varied in an attempt to optimize biomass growth and oil yield. Oil production in EDCCs reached a maximum of 5% (w/w) of total lipids. Although much lower than lipid production in seed, lipid profiles of EDCCs remain identical to those produced in planta. The expression levels of five major transcription factors (TFs), as well as KAS1, accA, DGAT1/2 and PDAT1 enzymes, and the OLE1 protein, all key components of the lipid biosynthesis pathway were also measured. Significant expression of LEC1, FUS3, ABI3, and WRI1 was found in endosperm cells throughout seed development, suggesting similar functions to their counterparts in Arabidopsis and providing a reference expression level for cell cultures. J. curcas EDCCs showed lower expression of most TFs compared with endosperm tissue, with the exception of WRI1 which had comparable expression levels in the two systems. Conversely, the enzymes KAS1, accA, and DGAT had the same or higher expression levels in EDCCs versus endosperm cells. Interestingly, the genes that encoded for DGAT1 and DGAT2 enzymes were found preferentially expressed in endosperm cells and EDCCs, respectively. Contrary to other studies, our findings indicate that the addition of ABA does not result in increased expression of genes involved in storage lipid biosynthesis.

MdKNOX19, a class II knotted-like transcription factor of apple, plays roles in ABA signalling/sensitivity by targeting ABI5 during organ development

The ABI5 transcription factor, which is a core component of the ABA signaling pathway, affects various plant processes, including seed development and germination and responses to environmental cues. The knotted1-like homeobox (KNOX) transcription factor has crucial functions related to plant development, including the regulation of various hormones. In this study, an ABA-responsive KNOX gene, MdKNOX19, was identified in apple (Malus domestica). The overexpression of MdKNOX19 increased the ABA sensitivity of apple calli, resulting in a dramatic up-regulation in the transcription of the Arabidopsis ABI5-like MdABI5 gene. Additionally, MdKNOX19 overexpression in Micro-Tom adversely affected fruit size and seed yield as well as enhanced ABA sensitivity and up-regulated SlABI5 transcription during seed germination and early seedling development. An examination of MdKNOX19-overexpressing Arabidopsis plants also revealed severe defects in seed development and up-regulated expression of ABA-responsive genes. Furthermore, we further confirmed that MdKNOX19 binds directly to the MdABI5 promoter to activate expression. Our findings suggest MdKNOX19 is a positive regulator of ABI5 expression, and the conserved module MdKNOX19-MdABI5-ABA may contribute to organ development.

A novel sucrose transporter gene IbSUT4 involves in plant growth and response to abiotic stress through the ABF-dependent ABA signaling pathway in Sweetpotato

BackgroundTo maintain sweetpotato (Ipomoea batatas (L.) Lam) growth and yield, sucrose must be transported from the leaves to the roots. Sucrose transporters or carriers (SUTs or SUCs) transport sucrose and are involved in plant growth and response to abiotic stress. However, the mechanisms of SUTs in sweetpotato abiotic stress resistance remains to be determined.ResultsIn the present study, we cloned a novel IbSUT4 gene; the protein encoded by this gene is localized in the tonoplast and plasma membrane. The plant growth was promoted in the IbSUT4 transgenic Arabidopsis thaliana lines, with increased expression of AtFT, a regulator of flowering time in plants. Over-expression of IbSUT4 in Arabidopsis thaliana resulted in higher sucrose content in the roots and lower sucrose content in the leaves, as compared to the wild-type (WT) plants, leading to improved stress tolerance during seedling growth. Moreover, we systematically analyzed the mechanisms of IbSUT4 in response to abiotic stress. The results suggest that the ABRE-motif was localized in the IbSUT4 promoter region, and the expression of the ABA signaling pathway genes (i.e., ABF2, ABF4, SnRK2.2, SnRK2.3, and PYL8/RCAR3) were induced, and the expression of ABI1 was inhibited.ConclusionsOur dates provide evidence that IbSUT4 is not only involved in plant growth but also is an important positive regulator in plant stress tolerance through the ABF-dependent ABA signaling pathway.

Involvement of abscisic acid, ABI5, and PPC2 in plant acclimation to low CO2.

Phosphoenolpyruvate carboxylase (PEPC) plays a pivotal role in the photosynthetic CO2 fixation of C4 plants. However, the functions of PEPCs in C3 plants are less well characterized, particularly in relation to low atmospheric CO2 levels. Of the four genes encoding PEPC in Arabidopsis, PPC2 is considered as the major leaf PEPC gene. Here we show that the ppc2 mutants suffered a growth arrest when transferred to low atmospheric CO2 conditions, together with decreases in the maximum efficiency of PSII (Fv/Fm) and lower levels of leaf abscisic acid (ABA) and carbohydrates. The application of sucrose, malate, or ABA greatly rescued the growth of ppc2 lines under low CO2 conditions. Metabolite profiling analysis revealed that the levels of glycine and serine were increased in ppc2 leaves, while the abundance of photosynthetic metabolites was decreased under these conditions. The transcript levels of encoding enzymes involved in glycine or serine metabolism was decreased in ppc2 in an ABI5-dependent manner. Like the ppc2 mutants, abi5-1 mutants had lower photosynthetic rates and Fv/Fm compared with the wild type under photorespiratory conditions (i.e. low CO2 availability). However, the growth of these mutants was similar to that of the wild type under non-photorespiratory (low O2) conditions. The constitutive expression of ABI5 prevented the growth arrest of ppc2 lines under low CO2 conditions. These findings demonstrate that PPC2 plays an important role in the acclimation of Arabidopsis plants to low CO2 availability by linking photorespiratory metabolism to primary metabolism, and that this is mediated, at least in part, through ABA- and ABI5-dependent processes.

PHYTOCHROME-INTERACTING FACTORS Interact with the ABA Receptors PYL8 and PYL9 to Orchestrate ABA Signaling in Darkness

PHYTOCHROME-INTERACTING FACTORS (PIFs) are a group of basic helix-loop-helix transcription factors that can physically interact with photoreceptors, including phytochromes and cryptochromes. It was previously demonstrated that PIFs accumulated in darkness and repressed seedling photomorphogenesis, and that PIFs linked different photosensory and hormonal pathways to control plant growth and development. In this study, we show that PIFs positively regulate the ABA signaling pathway during the seedling stage specifically in darkness. We found that PIFs positively regulate ABI5 transcript and protein levels in darkness in response to exogenous ABA treatment by binding directly to the G-box motifs in the ABI5 promoter. Consistently, PIFs and the G-box motifs in the ABI5 promoter determine ABI5 expression in darkness, and overexpression of ABI5 could rescue the ABA-insensitive phenotypes of pifq mutants in the dark. Moreover, we discovered that PIFs can physically interact with the ABA receptors PYL8 and PYL9, and that this interaction is not regulated by ABA. Further analyses showed that PYL8 and PYL9 promote PIF4 protein accumulation in the dark and enhance PIF4 binding to the ABI5 promoter, but negatively regulate PIF4-mediated ABI5 activation. Taken together, our data demonstrate that PIFs interact with ABA receptors to orchestrate ABA signaling in darkness by controlling ABI5 expression, providing new insights into the pivotal roles of PIFs as signal integrators in regulating plant growth and development.

Regulation of ABI5 expression by ABF3 during salt stress responses in Arabidopsis thaliana

BackgroundBasic region/leucine zippers (bZIPs) are transcription factors (TFs) encoded by a large gene family in plants. ABF3 and ABI5 are Group A bZIP TFs that are known to be important in abscisic acid (ABA) signaling. However, questions of whether ABF3 regulates ABI5 are still present.ResultsIn vitro kinase assay results showed that Thr-128, Ser-134, and Thr-451 of ABF3 are calcium-dependent protein kinase phosphorylation sites. Bimolecular fluorescence complementation (BiFC) analysis results showed a physical interaction between ABF3 and 14-3-3ω. A Thr-451 to Ala point mutation abolished the interaction but did not change the subcellular localization. In addition, the Arabidopsis protoplast transactivation assay using a luciferase reporter exhibited ABI5 activation by either ABF3 alone or by co-expression of ABF3 and 14-3-3ω. Moreover, chromatin immunoprecipitation-qPCR results showed that in Arabidopsis, ABI5 ABA-responsive element binding proteins (ABREs) of the promoter region (between − 1376 and − 455) were enriched by ABF3 binding under normal and 150 mM NaCl salt stress conditions.ConclusionTaken together, our results demonstrated that ABI5 expression is regulated by ABF3, which could contribute to salt stress tolerance in Arabidopsis thaliana.

Correction: Arabidopsis WRKY6 Transcription Factor Acts as a Positive Regulator of Abscisic Acid Signaling during Seed Germination and Early Seedling Development.

[This corrects the article DOI: 10.1371/journal.pgen.1005833.].

Arabidopsis small ubiquitin-related modifier protease ASP1 positively regulates abscisic acid signaling during early seedling development.

The small ubiquitin-related modifier (SUMO) modification plays an important role in the regulation of abscisic acid (ABA) signaling, but the function of the SUMO protease, in ABA signaling, remains largely unknown. Here, we show that the SUMO protease, ASP1 positively regulates ABA signaling. Mutations in ASP1 resulted in an ABA-insensitive phenotype, during early seedling development. Wild-type ASP1 successfully rescued, whereas an ASP1 mutant (C577S), defective in SUMO protease activity, failed to rescue, the ABA-insensitive phenotype of asp1-1. Expression of ABI5 and MYB30 target genes was attenuated in asp1-1 and our genetic analyses revealed that ASP1 may function upstream of ABI5 and MYB30. Interestingly, ASP1 accumulated upon ABA treatment, and ABA-induced accumulation of ABI5 (a positive regulator of ABA signaling) was abolished, whereas ABA-induced accumulation of MYB30 (a negative regulator of ABA signaling) was increased in asp1-1. These findings support the hypothesis that increased levels of ASP1, upon ABA treatment, tilt the balance between ABI5 and MYB30 towards ABI5-mediated ABA signaling.

MiR-181a/b induce the growth, invasion, and metastasis of neuroblastoma cells through targeting ABI1.

Neuroblastoma is a pediatric malignancy, and the clinical phenotypes range from localized tumors with excellent outcomes to widely metastatic disease in which long-term survival is approximately 40%, despite intensive therapy. Emerging evidence suggests that aberrant miRNA regulation plays a role in neuroblastoma, but the miRNA functions and mechanisms remain unknown. miR-181 family members were detected in 32 neuroblastoma patients, and the effects of miR-181a/b on cell viability, invasion, and migration were evaluated in vitro and in vivo. A parallel global mRNA expression profile was obtained for neuroblastoma cells overexpressing miR-181a. The potential targets of miR-181a/b were validated. miR-181a/b expression levels were positively associated with MYCN amplification and neuroblastoma aggressiveness. Moreover, ectopic miR-181a/b expression significantly induced the growth and invasion of neuroblastoma cells in vitro and in vivo. Microarray analysis revealed that mRNAs were consistently downregulated after miR-181a overexpression, leading to cell migration. In addition, the expression of ABI1 was suppressed by miR-181a/b, and ABI1 was validated as a direct target of miR-181a/b. We concluded that miR-181a/b were significantly upregulated in aggressive neuroblastoma, which enhanced its tumorigenesis and progression by suppressing the expression of ABI1.

HRB2 and BBX21 interaction modulates Arabidopsis ABI5 locus and stomatal aperture.

Blue light triggers the opening of stomata in the morning to allow CO2 uptake and water loss through transpiration. During the day, plants may experience periodic drought and accumulate abscisic acid (ABA). ABA antagonizes blue light signalling through phosphatidic acid and reduces stomatal aperture. This study reveals a molecular mechanism by which two light signalling proteins interact to repress ABA signalling in the control of stomatal aperture. A hypersensitive to red and blue 2 (hrb2) mutant has a defective ATP-dependent chromatin-remodelling factor, PKL, in the chromodomain/helicase/DNA binding family. HRB2 enhances the light-induced expression of a B-box transcription factor gene, BBX21. BBX21 binds a T/G box in the ABI5 promoter and recruits HRB2 to modulate the chromatin structure at the ABI5 locus. Mutation in either HRB2 or BBX21 led to reduced water loss and ABA hypersensitivity. This hypersensitivity to ABA was well explained by the enhanced expression of the ABA signalling gene ABI5 in both mutants. Indeed, stomatal aperture was significantly reduced by ABI5 overexpression in the absence or presence of ABA under monochromatic light conditions. Overall, we present a regulatory loop in which two light signalling proteins repress ABA signalling to sustain gas exchange when plants experience periodic drought.

Cross-talk between ABA and sugar signaling is mediated by the ACGT core and CE1 element reciprocally in OsTIP3;1 promoter

Recently, much effort has been made to determine the molecular links and cross-talk between sugar and abscisic acid (ABA) signaling pathways. ABA-inducible expression of OsTIP3;1, encoding a rice tonoplast intrinsic protein, was enhanced by sugar depletion. Such a stimulatory increase in OsTIP3;1 expression under sugar-starvation is possibly not owing to changes in endogenous ABA content. The transient expression assay indicated that the 5′ flanking region of OsTIP3;1 delivered similar collaborative responsiveness to starvation and ABA, suggesting that this gene promoter could be a good molecular probe to examine the interaction between sugar and ABA signaling pathways. Targeted mutagenesis demonstrated that disruption of ACGT cores decreased the induction of OsTIP3;1 promoter activity under either starvation or ABA, whereas mutation of coupling element 1 (CE1), which is an ABI4 binding site, reversely increased it, suggesting that those two distinct cis-regulatory elements reciprocally regulate the responsiveness of this promoter to both sugar and ABA. Consistent with this result, antisense inhibition of ABI4 increased the OsTIP3;1 promoter activity. ABI4 expression was also enhanced by sugars and repressed by ABA, suggesting that reduced ABI4 binding to CE1 in the absence of sugar and presence of ABA could increase ABA-induction of the OsTIP3;1 promoter activity.

Glutamate Receptor Homolog3.4 is Involved in Regulation of Seed Germination Under Salt Stress in Arabidopsis.

Seed germination is sensitive to salt stress. ABA and Ca2+ are involved in the regulation of seed germination under salt stress. Ca2+ influx mediated by glutamate receptors (GLRs) plays important roles in many physiological processes in plants. Here, we investigated the correlation of GLRs, Ca2+ and ABA during seed germination in response to salt stress by using Arabidopsis thaliana wild-type and T-DNA insertion knockout mutants of glutamate receptor homolog3.4. We demonstrated that atglr3.4-1 and atglr3.4-2 mutants were more sensitive to NaCl during seed germination and post-germination growth than wild-type plants. Treatments of wild-type seedlings with NaCl evoked a marked elevation in cytosolic Ca2+ activity ([Ca2+]cyt), and the elevation was inhibited by antagonists of GLRs, while the NaCl-induced elevation in [Ca2+]cyt was impaired in atglr3.4-1 and atglr3.4-2 mutants. Moreover, the mutants exhibited a lower expression of SOS3, SOS2 and SOS1, and greater accumulation of Na+ than wild-type seeds in the presence of NaCl. Mutation of AtGLR3.4 rendered the mutants more sensitive to ABA, while overexpression of AtGLR3.4 made the transgenic lines more tolerant to ABA in terms of seed germination. However, there was no difference in ABA content between atglr3.4 mutants and wild-type seeds, accompanied by lower expression of ABI3 and ABI4 in atglr3.4 mutants when challenged with NaCl. These results demonstrate that AtGLR3.4-mediated Ca2+ influx may be involved in the regulation of seed germination under salt stress by modulating Na+ accumulation through the SOS pathway.

Arabidopsis plants exposed to dioxin result in a WRINKLED seed phenotype due to 20S proteasomal degradation of WRI1.

Dioxins are highly toxic persistent organic pollutants bioaccumulated by both plants and animals that cause severe developmental abnormalities in humans. We investigated the effects of dioxins on seed development in Arabidopsis. Plants were exposed to various concentrations of the most toxic congener of dioxins, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and the effects on seed development were analysed in-depth at transcriptome, proteome and metabolome levels. Exposure to dioxin led to generalized effects on vegetative tissues plus a specific set of perturbations to seed development. Mature seeds from TCDD-treated plants had a characteristic 'wrinkled' phenotype, due to a two-thirds reduction in storage oil content. Transcriptional analysis of a panel of genes related to lipid and carbohydrate metabolism was consistent with the observed biochemical phenotypes. There were increases in WRI1 and LEC1 expression but decreases in ABI3 and FUS3 expression, which is puzzling in view of the low seed oil phenotype. This anomaly was explained by increased expression of 20S proteasome components that resulted in a substantial degradation of WRI1 protein, despite the up-regulation of the WRI1 gene. Our findings reveal novel effects of dioxins that lead to altered gene regulation patterns that profoundly affect seed development in Arabidopsis.

ABI3, a component of the WAVE2 complex, is potentially regulated by PI3K/AKT pathway.

We previously reported that ABI3 expression is lost in follicular thyroid carcinomas and its restoration significantly inhibited cell growth, invasiveness, migration, and reduced tumor growth in vivo. The mechanistic basis by which ABI3 exerts its tumor suppressive effects is not fully understood. In this study, we show that ABI3 is a phosphoprotein. Using proteomic array analysis, we showed that ABI3 modulated distinct cancer-related pathways in thyroid cancer cells. The KEA analysis found that PI3K substrates were enriched and forced expression of ABI3 markedly decreased the phosphorylation of AKT and the downstream-targeted protein pGSK3β. We next used immunoprecipitation combined with mass spectrometry to identify ABI3-interacting proteins that may be involved in modulating/integrating signaling pathways. We identified 37 ABI3 partners, including several components of the canonical WAVE regulatory complex (WRC) such as WAVE2/CYF1P1/NAP1, suggesting that ABI3 function might be regulated through WRC. Both, pharmacological inhibition of the PI3K/AKT pathway and mutation at residue S342 of ABI3, which is predicted to be phosphorylated by AKT, provided evidences that the non-phosphorylated form of ABI3 is preferentially present in the WRC protein complex. Collectively, our findings suggest that ABI3 might be a downstream mediator of the PI3K/AKT pathway that might disrupt WRC via ABI3 phosphorylation.

Artificial targeting of a nucleus-encoded RNA-binding protein AtRZ1a to chloroplasts affects flowering and ABA response of Arabidopsis thaliana

Despite the fact that many nucleus-encoded RNA-binding proteins (RBPs) are targeted to chloroplasts and play essential roles in RNA metabolism in chloroplasts, the question of whether artificial targeting of a nucleus-encoded RBP to chloroplasts affects chloroplast function and plant growth has never been addressed. In this study, a nuclear zinc finger-containing Arabidopsis RBP, designated AtRZ1a, which was previously shown to play a role in stress response, was artificially targeted to chloroplasts, and the growth and stress response of the transgenic plants were evaluated. Confocal analysis of the cellular localization of the cTP_AtRZ1a protein containing the N-terminal chloroplast transit peptide (cTP) from rubisco small subunit revealed that the cTP_AtRZ1a fusion protein is successfully targeted to chloroplasts. When grown under normal conditions, flowering of the transgenic plants was delayed, and the FLC expression was significantly upregulated in the transgenic plants. Artificial targeting of AtRZ1a to chloroplasts severely inhibited seedling growth of the plants in the presence of ABA by upregulating expression of ABA signaling-related genes ABI3 and ABI4. Taken together, these results suggest that artificial targeting of a nucleus-encoded AtRZ1a to chloroplasts affects the growth and development of Arabidopsis under normal or ABA treated conditions.

Abstract P6-01-14: Dissection of the Abl interactor 1 signaling in metastatic breast cancer cells

Abstract Abl interactor 1 (Abi1) is a key component of WAVE regulatory complex (WRC) that regulates actin cytoskeleton reorganization, membrane receptor signaling, and intracellular trafficking. Recent in vitro and in vivo studies as well as the studies of human patient samples suggest that Abi1 may play an important role in breast cancer (BC) metastasis. To determine if Abi1 is deregulated in metastatic breast cancer cells, we compared the expression of Abi1 in a panel of human breast cancer cell lines exhibiting distinct metastatic properties. In comparison with low-metastatic ER+/PR+ BC lines MCF-7 and T47D and a non-transforming mammary epithelial cell line MCF-10A, two ER-BC lines CN34 and MDA 231 and their sublines selected by high metastatic potential to bone (231Bo), brain (231Br and CN34Br), and lung (231LM2) show significantly reduced Abi1 protein levels, with greater reduction found in highly metastatic sublines. It is unlikely that the down-regulation of Abi1 expression in these metastatic BC cells occurs at mRNA level, because RT-PCR analysis shows no significant difference in Abi1 mRNA levels among the cells analyzed. Interestingly, despite a reduction in protein level, the tyrosine phosphorylation of Abi1 was increased in CN34, MDA231, and their metastatic sublines compared to low-metastatic BC cells. The tyrosine phosphorylation of Abi1 is stimulated by epidermal growth factor (EGF) in these cells and Abi1 is recruited to EGF receptor (EGFR) upon EGF stimulation. The deregulation of Abi1 in metastatic BC cells is mediated by Abl tyrosine kinases, as the inhibition of Abl family kinases by imatinib not only attenuates EGF-stimulated Abi1 tyrosine phosphorylation but also rescue the down-regulation of Abi1 protein in metastatic BC cells. These findings are consistent with a role of the EGFR-Abl-Abi1 signaling in BC metastasis. The studies suggest that this pathway may serve as a biomarker for diagnosis and/or prognosis of metastatic breast cancer. We have identified the tyrosine 213 in Abi1 as a major phosphorylation site stimulated by Abl kinases and have generated a unique conditional Abi1 depletion/re-expression system to dissect how the deregulation of the Abi1 signaling contributes to BC metastasis. Citation Format: Jiang P, Tang H, Hogan H, Tate S, Ryan W, Paul L, Liu X, Zonghan D. Dissection of the Abl interactor 1 signaling in metastatic breast cancer cells [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P6-01-14.

A WD40 protein, AtGHS40, negatively modulates abscisic acid degrading and signaling genes during seedling growth under high glucose conditions.

The Arabidopsis thaliana T-DNA insertion mutant glucose hypersensitive (ghs) 40-1 exhibited hypersensitivity to glucose (Glc) and abscisic acid (ABA). The ghs40-1 mutant displayed severely impaired cotyledon greening and expansion and showed enhanced reduction in hypocotyl elongation of dark-grown seedlings when grown in Glc concentrations higher than 3%. The Glc-hypersensitivity of ghs40-1 was correlated with the hyposensitive phenotype of 35S::AtGHS40 seedlings. The phenotypes of ghs40-1 were recovered by complementation with 35S::AtGHS40. The AtGHS40 (At5g11240) gene encodes a WD40 protein localized primarily in the nucleus and nucleolus using transient expression of AtGHS40-mRFP in onion cells and of AtGHS40-EGFP and EGFP-AtGHS40 in Arabidopsis protoplasts. The ABA biosynthesis inhibitor fluridone extensively rescued Glc-mediated growth arrest. Quantitative real time-PCR analysis showed that AtGHS40 was involved in the control of Glc-responsive genes. AtGHS40 acts downstream of HXK1 and is activated by ABI4 while ABI4 expression is negatively modulated by AtGHS40 in the Glc signaling network. However, AtGHS40 may not affect ABI1 and SnRK2.6 gene expression. Given that AtGHS40 inhibited ABA degrading and signaling gene expression levels under high Glc conditions, a new circuit of fine-tuning modulation by which ABA and ABA signaling gene expression are modulated in balance, occurred in plants. Thus, AtGHS40 may play a role in ABA-mediated Glc signaling during early seedling development. The biochemical function of AtGHS40 is also discussed.

Arabidopsis YL1/BPG2 Is Involved in Seedling Shoot Response to Salt Stress through ABI4.

The chloroplast-localized proteins play roles in plant salt stress response, but their mechanisms remain largely unknown. In this study, we screened a yellow leaf mutant, yl1-1, whose shoots exhibited hypersensitivity to salt stress. We mapped YL1 to AT3G57180, which encodes a YqeH-type GTPase. YL1, as a chloroplast stroma-localized protein, could be markedly reduced by high salinity. Upon exposure to high salinity, seedling shoots of yl1-1 and yl1-2 accumulated significantly higher levels of Na+ than wild type. Expression analysis of factors involved in plant salt stress response showed that the expression of ABI4 was increased and HKT1 was evidently suppressed in mutant shoots compared with the wild type under normal growth conditions. Moreover, salinity effects on ABI4 and HKT1 were clearly weakened in the mutant shoots, suggesting that the loss of YL1 function impairs ABI4 and HKT1 expression. Notably, the shoots of yl1-2 abi4 double mutant exhibited stronger resistance to salt stress and accumulated less Na+ levels after salt treatment compared with the yl1-2 single mutant, suggesting the salt-sensitive phenotype of yl1-2 seedlings could be rescued via loss of ABI4 function. These results reveal that YL1 is involved in the salt stress response of seedling shoots through ABI4.