RESUMEN
KEY MESSAGE: The chloroplast-localized protein CSAP is an ABA-responsive factor and positively regulates dark-induced senescence. This phenomenon is controlled by SAUL1 in Arabidopsis. We report here that CSAP (Chloroplast-localized Senescence-Associated Protein, AT5G39520) functions as a positive regulator of senescence and is controlled by SAUL1 (Senescence Associated E3 Ubiquitin Ligase 1) in Arabidopsis. CSAP transcript level was gradually increased when senescence was progressed. Under dark conditions, the csap mutant showed delayed leaf senescence and reduced chlorophyll breakdown, but overexpression of CSAP accelerated leaf senescence and expressions of chlorophyll catabolic genes were up-regulated compared to the wild-type (WT). NCED3 and AAO3, which are involved in ABA biosynthesis, also showed higher expression in the overexpression lines than the WT. It is known that the CSAP transcript is increased in the saul1 mutant that shows precocious senescence. In our experiments, we confirmed that CSAP interacts with SAUL1 by the yeast two-hybrid and pull-down assays. In addition, we found that SAUL1 decreases the stability of CSAP in the presence of ABA. Taken together, we suggest that CSAP accelerates leaf senescence in the dark and this process is controlled by SAUL1.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de Cloroplastos/metabolismo , Oscuridad , Proteínas de la Membrana/metabolismo , Hojas de la Planta/crecimiento & desarrollo , Ubiquitina-Proteína Ligasas/metabolismo , Ácido Abscísico/farmacología , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Cloroplastos/genética , Cloroplastos/efectos de los fármacos , Cloroplastos/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Proteínas de la Membrana/genética , Mutación/genética , Fenotipo , Hojas de la Planta/efectos de los fármacos , Plantas Modificadas Genéticamente , Unión Proteica/efectos de los fármacos , Estabilidad Proteica/efectos de los fármacosRESUMEN
KEY MESSAGE: PpCKX1 localizes to vacuoles and is dominantly expressed in the stem cells. PpCKX1 regulates developmental changes with increased growth of the rhizoid and enhances dehydration and salt tolerance. Cytokinins (CKs) are plant hormones that regulate plant development as well as many physiological processes, such as cell division, leaf senescence, control of shoot/root ratio, and reproductive competence. Cytokinin oxidases/dehydrogenases (CKXs) control CK concentrations by degradation, and thereby influence plant growth and development. In the moss Physcomitrella patens, an evolutionarily early divergent plant, we identified six putative CKXs that, by phylogenetic analysis, form a monophyletic clade. We also observed that ProPpCKX1:GUS is expressed specifically in the stem cells and surrounding cells and that CKX1 localizes to vacuoles, as indicated by Pro35S:PpCKX1-smGFP. Under normal growth conditions, overexpression of PpCKX1 caused many phenotypic changes at different developmental stages, and we suspected that increased growth of the rhizoid could affect those changes. In addition, we present evidence that the PpCKX1-overexpressor plants show enhanced dehydration and salt stress tolerance. Taken together, we suggest that PpCKX1 plays regulatory roles in development and adaptation to abiotic stresses in this evolutionarily early land plant species.
Asunto(s)
Bryopsida/enzimología , Bryopsida/crecimiento & desarrollo , Oxidorreductasas/metabolismo , Tolerancia a la Sal , Bryopsida/genética , Citocininas/metabolismo , Deshidratación , Regulación de la Expresión Génica de las Plantas , Fenotipo , Filogenia , Plantas Modificadas Genéticamente , Estrés Salino/genética , Tolerancia a la Sal/genética , Células Madre/metabolismo , Vacuolas/metabolismoRESUMEN
Nucleases are a very diverse group of enzymes that play important roles in many crucial physiological processes in plants. We previously reported that the highly conserved region (HCR), domain of unknown function 151 (DUF151) and UV responsive (UVR) domain-containing OmBBD is a novel nuclease that does not share homology with other well-studied plant nucleases. Here, we report that DUF151 domain-containing proteins are present in bacteria, archaea and only Viridiplantae kingdom of eukarya, but not in any other eukaryotes. Two Arabidopsis homologs of OmBBD, AtBBD1 and AtBBD2, shared 43.69% and 44.38% sequence identity and contained all three distinct domains of OmBBD. We confirmed that the recombinant MBP-AtBBD1 and MBP-AtBBD2 exhibited non-substrate-specific DNase and RNase activity, like OmBBD. We also found that a metal cofactor is not necessarily required for DNase activity of AtBBD1 and AtBBD2, but their activities were much enhanced in the presence of Mg2+ or Mn2+. Using a yeast two-hybrid assay, we found that AtBBD1 and AtBBD2 each form a homodimer but not a heterodimer and that the HCR domain is possibly crucial for dimerization.
Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/enzimología , Arabidopsis/genética , Endonucleasas/metabolismo , Dominios Proteicos/genética , Secuencia de Aminoácidos , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Archaea/genética , Archaea/metabolismo , Bacterias/genética , Chlamydomonas reinhardtii/genética , Desoxirribonucleasas/genética , Desoxirribonucleasas/metabolismo , Endonucleasas/genética , Eucariontes/genética , Eucariontes/metabolismo , Evolución Molecular , Magnesio/química , Manganeso/química , Oryza/enzimología , Oryza/genética , Oryza/metabolismo , Filogenia , Multimerización de Proteína/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Homología de Secuencia de AminoácidoRESUMEN
KEY MESSAGE: The trichome-related protein (TRP) is a novel transcription factor (TF) that negatively regulates trichome initiation-related TFs through gibberellin (GA) signaling. Trichomes, which are outgrowths of leaf epidermal cells, provide the plant with a first line of defense against damage from herbivores and reduce transpiration. The initiation and development of trichomes are regulated by a network of positively or negatively regulating transcription factors (TFs). However, little information is currently available on transcriptional regulation related to trichome formation. Here, we report a novel TF Trichome-Related Protein (TRP) that was observed to negatively regulate the trichome initiation-related TFs through gibberellic acid (GA) signaling. ProTRP:GUS revealed that TRP was only expressed in the trichome. The TRP loss-of-function mutant (trp) had an increased number of trichomes on the flower, cauline leaves, and main inflorescence stems compared to the wild-type. In contrast, TRP overexpression lines (TRP-Ox) exhibited a decreased number of trichomes on cauline leaves and main inflorescence stem following treatment with exogenous GA. Moreover, the expressions of trichome initiation regulators (GIS, GIS2, ZFP8, GL1, and GL3) increased in trp plants but decreased in TRP-Ox lines after GA treatment. TRP was observed to physically interact with ZFP5, a C2H2 TF that controls trichome cell development through GA signaling, both in vivo and in vitro. Based on these results, we suggest that TRP functions upstream of the trichome initiation regulators and represses the binding of ZFP5 to the ZFP8 promoter.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Giberelinas/farmacología , Canales de Potencial de Receptor Transitorio/metabolismo , Tricomas/metabolismo , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Mutación , Reguladores del Crecimiento de las Plantas/farmacología , Transducción de Señal/efectos de los fármacos , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Canales de Potencial de Receptor Transitorio/genéticaRESUMEN
Preventing or effectively treating metastatic uveal melanoma (UM) is critical because it occurs in about half of patients and confers a very poor prognosis. There is emerging evidence that hepatocyte growth factor (HGF) and insulin-like growth factor 1 (IGF-1) promote metastasis and contribute to the striking metastatic hepatotropism observed in UM metastasis. However, the molecular mechanisms by which HGF and IGF-1 promote UM liver metastasis have not been elucidated. ASAP1, which acts as an effector for the small GTPase ARF6, is highly expressed in the subset of uveal melanomas most likely to metastasize. Here, we found that HGF and IGF-1 hyperactivate ARF6, leading to its interaction with ASAP1, which then acts as an effector to induce nuclear localization and transcriptional activity of NFAT1. Inhibition of any component of this pathway impairs cellular invasiveness. Additionally, knocking down ASAP1 or inhibiting NFAT signaling reduces metastasis in a xenograft mouse model of UM. The discovery of this signaling pathway represents not only an advancement in our understanding of the biology of uveal melanoma metastasis but also identifies a novel pathway that could be targeted to treat or prevent metastatic uveal melanoma.
Asunto(s)
Melanoma , Neoplasias de la Úvea , Humanos , Animales , Ratones , Factor de Crecimiento de Hepatocito/genética , Factor de Crecimiento de Hepatocito/metabolismo , Factor I del Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/metabolismo , Melanoma/patología , Neoplasias de la Úvea/metabolismo , Modelos Animales de Enfermedad , Proteínas Adaptadoras Transductoras de Señales/metabolismoRESUMEN
Despite being toxic at a high concentrations, reactive oxygen species (ROS) play a pivotal role as signaling molecules in responses to stress and regulation of plant development. The mitochondrial electron transport chain (ETC) is the major source of ROS in cells. Although the regulation of ROS in mitochondria has been well elucidated, the protein-protein interaction-based regulation of ETC members has not been well elucidated. In this study, we identified a CBS domain-containing protein, CBSX3, and found that CBSX3 activates o-type thioredoxin (Trx-o2) in mitochondria. In addition, we found that Trx-o2 interacts with SDH1, a subunit of ETC complex II. Knockdown (KD) of CBSX3 revealed anther indehiscence due to deficient lignin deposition caused by insufficient ROS accumulation, and increased expression of genes related to cell cycle and accelerated plant growth. However, in the CBSX3-overexpression plants, ROS accumulation increased, and cell cycle-related gene expression decreased, and thereby plant growth was retarded and leaf size decreased. Moreover, KD of CBSX3 and Trx-o2 conferred resistance to mitochondria ETC inhibitors in terms of ROS release. Taken together, we suggest that CBSX3-Trx-o2 is a ROS generation regulator of mitochondria in plants and plays an important role in regulating plant development and the redox system.
Asunto(s)
Arabidopsis/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Mitocondrias/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Succinato Deshidrogenasa/metabolismo , Tiorredoxinas/metabolismoRESUMEN
Plant thioredoxins (Trxs) act as antioxidants and function as redox regulators in the chloroplast. Although the regulation of ROS in chloroplasts is well elucidated, the precise regulation mechanism of Trx remains unknown. Here, we characterize a novel chloroplast protein, Lon domain-containing protein 1 (LCP1), which contains only a Lon domain, the precise function of which is not known. We find that LCP1 interacts with Trx-y2 and represses its activity, and that knockdown (KD) of LCP1 causes anther indehiscence due to deficient lignin deposition. In addition, LCP1 KD plants show less ROS accumulation and lower expression of ROS-responsive marker genes than the wild-type plant. Taken together, we suggest that LCP1 directly regulates Trx-y2 and controls H2 O2 levels and, thereby, regulates lignin polymerization in the anther endothecium.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Flores/metabolismo , Regulación de la Expresión Génica de las Plantas , Tiorredoxinas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Flores/genética , Peróxido de Hidrógeno/metabolismo , Lignina/biosíntesis , Lignina/genética , Tiorredoxinas/genéticaRESUMEN
Osmotic stress, caused by drought, salinity, or PEG (polyethylene glycol), is one of the most important abiotic factors that hinder plant growth and development. In Arabidopsis, more than 100 R2R3-MYB transcription factors (TFs) have been identified, and many of them are involved in the transcriptional regulation of a variety of biological processes related to growth and development, as well as responses to biotic and abiotic stresses. However, the MYB TF involving in both plant development and stress response has rarely been reported. We report here that Arabidopsis AtMYB109, a R2R3-MYB TF, functions as a negative regulator of stomatal closure under osmotic stress as well as of pollen tube elongation. Under PEG-induced osmotic stress, whole leaves of AtMYB109-OXs were intensely wilted, while leaves of the wild-type (WT) and myb109 were weakly affected. Moreover, we confirmed that the wilting in AtMYB109-OXs was more severe than in WT and myb109 under drought conditions, and that after re-watering, WT and myb109 plants promptly recovered, while AtMYB109-OXs failed to survive. In addition, stomatal closure was delayed in the AtMYB109-OXs compared to the WT and myb109. However, proline content and the expression of stress-induced and proline synthesis genes were higher in the overexpression lines than in WT and myb109. Then, we observed that the expression of ICS1, a key gene in SA biosynthesis, was greatly suppressed in AtMYB109-OXs. In addition, we found that AtMYB109 expression gradually increased until the flowers were fully opened and thereafter dramatically decreased during silique development. The pollen tube growth was significantly suppressed in AtMYB109-OXs compared to the WT and myb109. Using EMSA and ChIP-qPCR, we confirmed that AtMYB109 bound to the promoter of RABA4D, a gene encoding a pollen development regulator. Taken together, we suggest the delayed stomatal closing and vulnerable phenotypes in the AtMYB109-OXs under osmotic stress are possibly directly or indirectly associated with a SA-mediated mechanism, and that AtMYB109 suppresses RABA4D that modulates pollen tube growth.