RESUMO
The FUSCA3 (FUS3) transcription factor is considered a master regulator of seed maturation because a wide range of seed maturation events are impaired in its defective mutant. To identify comprehensively genes under the control of FUS3, two types of microarray experiments were performed. First, transgenic plants in which FUS3 expression could be induced by the application of estrogen (ESTR) were used to identify any genes up-regulated in young seedlings of Arabidopsis in response to the ectopic expression of FUS3. Secondly, the transcriptomes of the fus3 mutant and wild-type developing seeds were compared. The combined results of these experiments identified genes under the relatively immediate and robust control of FUS3 during seed development. The analysis has extended the range of identified gene types under the control of FUS3. The genes positively controlled by FUS3 are not confined to previously known seed maturation-related genes and include those involved in the production of secondary metabolites, such as glucosinolates, phenylpropanoids and flavonoids, and those involved in primary metabolism, such as photosynthesis and fatty acid biosynthesis. Furthermore, several different patterns were identified in the manner of ectopic activation by FUS3 with respect to the induction kinetics and ABA requirement of downstream gene induction depending on the nature of developmental regulation, suggesting mechanistic diversity of gene regulation by FUS3.
Assuntos
Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Sementes/crescimento & desenvolvimento , Sementes/genética , Fatores de Transcrição/genética , Ácido Abscísico/farmacologia , Alelos , Arabidopsis/efeitos dos fármacos , Arabidopsis/metabolismo , Estrogênios/farmacologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , RNA de Plantas , Plântula/genética , Plântula/metabolismo , Sementes/metabolismo , Fatores de Transcrição/metabolismo , Ativação Transcricional , Regulação para CimaRESUMO
Arabidopsis ABSCISIC ACID INSENSEITIVE3 (ABI3), FUSCA3 (FUS3) and LEAFY COTYLEDON1 (LEC1) encode key transcription factors that control seed maturation events, including seed storage protein (SSP) accumulation. Although lec1 mutations are known to down-regulate SSP gene expression as the fus3 or abi3 mutation does, the mechanisms by which LEC1 regulates SSP gene expression are largely unknown compared with the mechanisms utilized by FUS3 or ABI3. We expressed LEC1 ectopically in transgenic plants using an artificial dexamethasone (Dex) induction system. The ectopic expression of LEC1 also resulted in the induction of FUS3 and ABI3 expression, which preceded the induction of SSP expression. The expression of FUS3 and ABI3 was found to be down-regulated in developing siliques of the lec1 mutant. Furthermore, the levels of ectopic SSP induction by LEC1 were greatly or moderately reduced in transgenic plants with an abi3 or fus3 mutant background, respectively. LEC1-induced ectopic expression of the At1g62290 aspartic protease gene, which was identified to be regulated preferentially by FUS3, was more severely affected in the fus3 mutant than in the abi3 mutant. From these data, we suggest that LEC1 controls the expression of the SSP genes in a hierarchical manner, which involves ABI3 and FUS3.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Sementes/crescimento & desenvolvimento , Fatores de Transcrição/metabolismo , Proteínas de Arabidopsis/genética , Ácido Aspártico Endopeptidases/genética , Ácido Aspártico Endopeptidases/metabolismo , Proteínas Estimuladoras de Ligação a CCAAT/genética , Dexametasona/farmacologia , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Plantas Geneticamente Modificadas , Fatores de Transcrição/genéticaRESUMO
The key transcription factors that control seed maturation, ABSCISIC ACID INSENSITIVE3 (ABI3) and FUSCA3 (FUS3), share homologous DNA-binding domains. Regulation of seed storage protein genes At2S3 and CRC by ABI3 and FUS3 was investigated using transgenic plants in which ABI3 and FUS3 could be ectopically induced by steroid hormones. Like ABI3, the presence of FUS3 led to expression of At2S3 and CRC in vegetative tissues. FUS3-mediated induction of CRC was completely dependent on exogenous abscisic acid (ABA), while At2S3 was weakly induced without ABA but strongly enhanced with ABA. This ABA dependency of FUS3-induced CRC and At2S3 expression was similar to that observed for ABI3. However, kinetic analysis revealed distinctions between the mechanisms of ABA-dependent CRC regulation by FUS3 or ABI3, and between target genes. While At2S3 activation by FUS3 was rapid, CRC induction by FUS3 in the presence of ABA, and by ABA followed by the presence of FUS3, took a significantly longer time (24-36 h). This suggested the involvement of an indirect mechanism requiring the ABA- and FUS3-dependent synthesis of intermediate regulatory factor(s). A chimeric protein composed of the FUS3 B3 domain, and a heterologous activation domain and nuclear localization signal exhibited a tight coupling with ABA regulation as observed for wild-type FUS3. Simultaneous induction of FUS3 and ABI3 did not result in the synergistic activation of CRC and At2S3. Based on these results, similarities and differences in the mechanisms of seed storage protein gene regulation by FUS3 and ABI3 are discussed.
