RESUMEN
Multiple mechanisms have been described for coordination of responses to the plant hormones auxin and brassinosteroids (Zhang et al., 2009). One unexplained phenomenon is the reliance of the auxin transcriptional response on a functional brassinosteroid pathway. In this study, we used luciferase reporters to interrogate the promoter of SMALL AUXIN-UP RNA15 (SAUR15), a well-characterized auxin and brassinosteroid early response gene in Arabidopsis (Arabidopsis thaliana). After identifying a minimal region sufficient for auxin response, we targeted predicted cis-regulatory elements contained within this sequence and found a critical subset required for hormone response. Specifically, reporter sensitivity to auxin treatment required two elements: a Hormone Up at Dawn (HUD)-type E-box and an AuxRE-related TGTCT element. Reporter response to brassinosteroid treatment relied on the same two elements. Consistent with these findings, the transcription factors BRASSINOSTEROID INSENSITIVE1-EMS SUPPESSOR1 and MONOPTEROS (MP)/ AUXIN RESPONSE FACTOR5 (ARF5) showed enhanced binding to the critical promoter region containing these elements. Treatment with auxin or brassinosteroids could enhance binding of either transcription factor, and brassinosteroid enhancement of MP/ARF5 binding required an intact HUD element. Conservation of clustered HUD elements and AuxRE-related sequences in promoters of putative SAUR15 orthologs in a number of flowering plant species, in combination with evidence for statistically significant clustering of these elements across all Arabidopsis promoters, provided further evidence of the functional importance of coordinated transcription factor binding.
Asunto(s)
Arabidopsis/genética , Arabidopsis/metabolismo , Ácidos Indolacéticos/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Secuencia de Bases , Brasinoesteroides/metabolismo , Secuencia Conservada , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Regiones Promotoras Genéticas , Secuencias Reguladoras de Ácidos Nucleicos , Elementos de Respuesta/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Plant form is shaped by a complex network of intrinsic and extrinsic signals. Light-directed growth of seedlings (photomorphogenesis) depends on the coordination of several hormone signals, including brassinosteroids (BRs) and auxin. Although the close relationship between BRs and auxin has been widely reported, the molecular mechanism for combinatorial control of shared target genes has remained elusive. Here we demonstrate that BRs synergistically increase seedling sensitivity to auxin and show that combined treatment with both hormones can increase the magnitude and duration of gene expression. Moreover, we describe a direct connection between the BR-regulated BIN2 kinase and ARF2, a member of the Auxin Response Factor family of transcriptional regulators. Phosphorylation by BIN2 results in loss of ARF2 DNA binding and repression activities. arf2 mutants are less sensitive to changes in endogenous BR levels, whereas a large proportion of genes affected in an arf2 background are returned to near wild-type levels by altering BR biosynthesis. Together, these data suggest a model where BIN2 increases expression of auxin-induced genes by directly inactivating repressor ARFs, leading to synergistic increases in transcription.
Asunto(s)
Proteínas de Arabidopsis/fisiología , Regulación de la Expresión Génica de las Plantas , Ácidos Indolacéticos/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/metabolismo , Proteínas Represoras/fisiología , Proteínas de Plantas/fisiología , Proteínas Quinasas/fisiología , Transcripción GenéticaRESUMEN
Recent studies suggest that hormones act through a web of interacting responses rather than through isolated linear pathways. This signal integration architecture may be one mechanism for increasing the specificity of outcomes in different cellular contexts. Several common themes for cross-regulation between pathways can be observed. Here, we propose a classification scheme for different levels of signaling pathway cross-regulation. This scheme is based on which parts of the individual pathways are acting as information conduits between pathways. Examples from the recent plant hormone biology literature are used to illustrate the different modes of interaction.
Asunto(s)
Reguladores del Crecimiento de las Plantas/fisiología , Fenómenos Fisiológicos de las Plantas , Ácido Abscísico/fisiología , Ciclopentanos/metabolismo , Giberelinas/metabolismo , Homeostasis , Ácidos Indolacéticos/metabolismo , Medicago/fisiología , Oxilipinas/metabolismo , Transducción de SeñalRESUMEN
We examined the chloroplast DNA (cpDNA) from plastids obtained from wild type maize (Zea mays L.) seedlings grown under different light conditions and from photosynthetic mutants grown under white light. The cpDNA was evaluated by real-time quantitative PCR, quantitative DNA fluorescence, and blot-hybridization following pulsed-field gel electrophoresis. The amount of DNA per plastid in light-grown seedlings declines greatly from stalk to leaf blade during proplastid-to-chloroplast development, and this decline is due to cpDNA degradation. In contrast, during proplastid-to-etioplast development in the dark, the cpDNA levels increase from the stalk to the blade. Our results suggest that DNA replication continues in the etioplasts of the upper regions of the stalk and in the leaves. The cpDNA level decreases rapidly, however, after dark-grown seedlings are transferred to light and the etioplasts develop into photosynthetically active chloroplasts. Light, therefore, triggers the degradation of DNA in maize chloroplasts. The cpDNA is retained in the leaf blade of seedlings grown under red, but not blue light. We suggest that light signaling pathways are involved in mediating cpDNA levels, and that red light promotes replication and inhibits degradation and blue light promotes degradation. For five of nine photosynthetic mutants, cpDNA levels in expanded leaves are higher than in wild type, indicating that nuclear genotype can affect the loss or retention of cpDNA.