RESUMEN
Auxin determines the developmental fate of plant tissues, and local auxin concentration is precisely controlled. The role of auxin transport in modulating local auxin concentration has been widely studied but the regulation of local auxin biosynthesis is less well understood. Here, we show that TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS (TAA1), a key enzyme in the auxin biosynthesis pathway in Arabidopsis thaliana is phosphorylated at Threonine 101 (T101). T101 phosphorylation status can act as an on/off switch to control TAA1-dependent auxin biosynthesis and is required for proper regulation of root meristem size and root hair development. This phosphosite is evolutionarily conserved suggesting post-translational regulation of auxin biosynthesis may be a general phenomenon. In addition, we show that auxin itself, in part via TRANS-MEMBRANE KINASE 4 (TMK4), can induce T101 phosphorylation of TAA1 suggesting a self-regulatory loop whereby local auxin signalling can suppress biosynthesis. We conclude that phosphorylation-dependent control of TAA1 enzymatic activity may contribute to regulation of auxin concentration in response to endogenous and/or external cues.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Ácidos Indolacéticos/metabolismo , Transducción de Señal/fisiología , Triptófano-Transaminasa/metabolismo , Proteínas de Arabidopsis/genética , Transporte Biológico/fisiología , Regulación de la Expresión Génica de las Plantas , Marchantia/metabolismo , Meristema/metabolismo , Simulación del Acoplamiento Molecular , Mutación , Fosforilación , Filogenia , Raíces de Plantas/metabolismo , Proteínas Serina-Treonina Quinasas , Triptófano-Transaminasa/química , Triptófano-Transaminasa/clasificaciónRESUMEN
Auxin is a fundamental plant hormone and its localization within organs plays pivotal roles in plant growth and development. Analysis of many Arabidopsis mutants that were defective in auxin biosynthesis revealed that the indole-3-pyruvic acid (IPA) pathway, catalyzed by the TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS (TAA) and YUCCA (YUC) families, is the major biosynthetic pathway of indole-3-acetic acid (IAA). In contrast, little information is known about the molecular mechanisms of auxin biosynthesis in rice. In this study, we identified a auxin-related rice mutant, fish bone (fib). FIB encodes an orthologue of TAA genes and loss of FIB function resulted in pleiotropic abnormal phenotypes, such as small leaves with large lamina joint angles, abnormal vascular development, small panicles, abnormal organ identity and defects in root development, together with a reduction in internal IAA levels. Moreover, we found that auxin sensitivity and polar transport activity were altered in the fib mutant. From these results, we suggest that FIB plays a pivotal role in IAA biosynthesis in rice and that auxin biosynthesis, transport and sensitivity are closely interrelated.
Asunto(s)
Ácidos Indolacéticos/metabolismo , Oryza/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/fisiología , Triptófano-Transaminasa/fisiología , Secuencia de Aminoácidos , Transporte Biológico/genética , Clonación Molecular , Datos de Secuencia Molecular , Oryza/enzimología , Oryza/metabolismo , Fenotipo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Alineación de Secuencia , Análisis de Secuencia de Proteína , Triptófano-Transaminasa/química , Triptófano-Transaminasa/genéticaRESUMEN
Malassezia yeasts are responsible for the widely distributed skin disease Pityriasis versicolor (PV), which is characterized by a hyper- or hypopigmentation of affected skin areas. For Malassezia furfur, it has been shown that pigment production relies on tryptophan metabolism. A tryptophan aminotransferase was found to catalyse the initial catalytic step in pigment formation in the model organism Ustilago maydis. Here, we describe the sequence determination, recombinant production and biochemical characterization of tryptophan aminotransferase MfTam1 from M. furfur. The enzyme catalyses the transamination from l-tryptophan to keto acids such as α-ketoglutarate with Km values for both substrates in the low millimolar range. Furthermore, MfTam1 presents a temperature optimum at 40°C and a pH optimum at 8.0. MfTam1 activity is highly dependent on pyridoxal phosphate (PLP), whereas compounds interfering with PLP, such as cycloserine (CS) and aminooxyacetate, inhibit the MfTam1 reaction. CS is known to reverse hyperpigmentation in PV. Thus, the results of the present study give a deeper insight into the role of MfTam1 in PV pathogenesis and as potential target for the development of novel PV therapeutics.
Asunto(s)
Indoles/química , Malassezia/enzimología , Piel/microbiología , Tiña Versicolor/microbiología , Triptófano-Transaminasa/química , Ácido Aminooxiacético/química , Clonación Molecular , Cicloserina/química , Escherichia coli/metabolismo , Proteínas Fúngicas/química , Humanos , Cetoácidos/química , Pigmentación , Pigmentos Biológicos/metabolismo , Fosfato de Piridoxal/química , Proteínas Recombinantes/química , Triptófano/químicaRESUMEN
Auxin is a plant growth regulator involved in diverse fundamental developmental responses. Much is now known about auxin transport, via influx and efflux carriers, and about auxin perception and its role in gene regulation. Many developmental processes are dependent on peaks of auxin concentration and, to date, attention has been directed at the role of polar auxin transport in generating and maintaining auxin gradients. However, surprisingly little attention has focussed on the role and significance of auxin biosynthesis, which should be expected to contribute to active auxin pools. Recent reports on the function of the YUCCA flavin monooxygenases and a tryptophan aminotransferase in Arabidopsis have caused us to look again at the importance of local biosynthesis in developmental processes. Many alternative and redundant pathways of auxin synthesis exist in many plants and it is emerging that they may function in response to environmental cues.
Asunto(s)
Ácidos Indolacéticos/metabolismo , Triptófano-Transaminasa/química , Arabidopsis/metabolismo , Ambiente , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Modelos Biológicos , Modelos Genéticos , Oxigenasas/química , Reguladores del Crecimiento de las Plantas/metabolismo , Raíces de Plantas/metabolismo , Triptófano/metabolismo , Yucca/metabolismoAsunto(s)
Arabidopsis/genética , Ácidos Indolacéticos/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Hormonas/genética , Hormonas/metabolismo , Ácidos Indolacéticos/química , Indoles/química , Indoles/metabolismo , Mutación , Fenotipo , Triptófano-Transaminasa/química , Triptófano-Transaminasa/metabolismoRESUMEN
Plants grown at high densities perceive a decrease in the red to far-red (R:FR) ratio of incoming light, resulting from absorption of red light by canopy leaves and reflection of far-red light from neighboring plants. These changes in light quality trigger a series of responses known collectively as the shade avoidance syndrome. During shade avoidance, stems elongate at the expense of leaf and storage organ expansion, branching is inhibited, and flowering is accelerated. We identified several loci in Arabidopsis, mutations in which lead to plants defective in multiple shade avoidance responses. Here we describe TAA1, an aminotransferase, and show that TAA1 catalyzes the formation of indole-3-pyruvic acid (IPA) from L-tryptophan (L-Trp), the first step in a previously proposed, but uncharacterized, auxin biosynthetic pathway. This pathway is rapidly deployed to synthesize auxin at the high levels required to initiate the multiple changes in body plan associated with shade avoidance.
Asunto(s)
Arabidopsis/fisiología , Ácidos Indolacéticos/metabolismo , Triptófano/metabolismo , Secuencia de Aminoácidos , Arabidopsis/genética , Vías Biosintéticas , Oscuridad , Regulación de la Expresión Génica de las Plantas , Datos de Secuencia Molecular , Hojas de la Planta/metabolismo , Alineación de Secuencia , Triptófano/biosíntesis , Triptófano-Transaminasa/química , Triptófano-Transaminasa/genética , Triptófano-Transaminasa/metabolismoRESUMEN
Plants have evolved a tremendous ability to respond to environmental changes by adapting their growth and development. The interaction between hormonal and developmental signals is a critical mechanism in the generation of this enormous plasticity. A good example is the response to the hormone ethylene that depends on tissue type, developmental stage, and environmental conditions. By characterizing the Arabidopsis wei8 mutant, we have found that a small family of genes mediates tissue-specific responses to ethylene. Biochemical studies revealed that WEI8 encodes a long-anticipated tryptophan aminotransferase, TAA1, in the essential, yet genetically uncharacterized, indole-3-pyruvic acid (IPA) branch of the auxin biosynthetic pathway. Analysis of TAA1 and its paralogues revealed a link between local auxin production, tissue-specific ethylene effects, and organ development. Thus, the IPA route of auxin production is key to generating robust auxin gradients in response to environmental and developmental cues.