The Arabidopsis HY5 gene has been defined genetically as a positive regulator of photomorphogenesis and recently has been shown to encode a basic leucine zipper type of transcription factor. Here, we report that HY5 is constitutively nuclear localized and is involved in light regulation of transcriptional activity of the promoters containing the G-box, a well-characterized light-responsive element (LRE). In vitro DNA binding studies suggested that HY5 can bind specifically to the G-box DNA sequences but not to any of the other LREs present in the light-responsive promoters examined. High-irradiance light activation of two synthetic promoters containing either the consensus G-box alone or the G-box combined with the GATA motif (another LRE) and the native Arabidopsis ribulose bisphosphate carboxylase small subunit gene RBCS-1A promoter, which has an essential copy of the G-box, was significantly compromised in the hy5 mutant. The hy5 mutation's effect on the high-irradiance light activation of gene expression was observed in both photosynthetic and nonphotosynthetic tissues. Furthermore, the characteristic phytochrome-mediated red light- and far-red light-reversible low-fluence induction of the G-box-containing promoters was diminished specifically in hy5 plants. These results suggest that HY5 may interact directly with the G-box in the promoters of light-inducible genes to mediate light-controlled transcriptional activity.

Higher plants are able to integrate environmental and endogenous signals to regulate gene expression for optimal development. To define the minimal sequence requirement sufficient to integrate light and developmental signals in controlling promoter activity, we carried out a systematic analysis of the roles of four well‐conserved ‘light‐responsive elements (LREs)’ common to many nuclear‐encoded photosynthetic genes. A gain‐of‐function assay using basal promoter‐reporter fusions in stable transgenic Arabidopsis was employed to demonstrate that pairwise combinations of the LREs, but not the individual elements alone, can confer light‐inducible expression to the reporter gene independently of the basal promoter context and the light‐triggered morphological changes. The activity of the synthetic promoters with the paired LREs can be modulated at least by the phytochrome system. Further, those synthetic light‐regulated promoters confer a photosynthetic cell‐specific expression pattern and respond to the chloroplast development state. Our data suggest that distinct combinatorial interactions of LREs can serve as minimal autonomous promoter determinants which integrate light and developmental signals and modulate promoter activity.

The Arabidopsis HY5 gene has been defined genetically as a positive regulator of photomorphogenesis and recently has been shown to encode a basic leucine zipper type of transcription factor. Here, we report that HY5 is constitutively nuclear localized and is involved in light regulation of transcriptional activity of the promoters containing the G-box, a well-characterized light-responsive element (LRE). In vitro DNA binding studies suggested that HY5 can bind specifically to the G-box DNA sequences but not to any of the other LREs present in the light-responsive promoters examined. High-irradiance light activation of two synthetic promoters containing either the consensus G-box alone or the G-box combined with the GATA motif (another LRE) and the native Arabidopsis ribulose bisphosphate carboxylase small subunit gene RBCS-1A promoter, which has an essential copy of the G-box, was significantly compromised in the hy5 mutant. The hy5 mutation's effect on the high-irradiance light activation of gene expression was observed in both photosynthetic and nonphotosynthetic tissues. Furthermore, the characteristic phytochrome-mediated red light- and far-red light-reversible low-fluence induction of the G-box-containing promoters was diminished specifically in hy5 plants. These results suggest that HY5 may interact directly with the G-box in the promoters of light-inducible genes to mediate light-controlled transcriptional activity.

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SummaryWe have studied the roles of PhyA, PhyB and CRY1 photoreceptors and the downstream light-signaling components, COP1 and DET1, in mediating high-irradiance light-controlled activity of promoters containing synthetic light-responsive elements (LRE). Promoters with paired LREs were able to respond to a wide spectrum of light through multiple photoreceptors, while the light-inducible single LRE promoters primarily responded to a specific wavelength of light. In addition, our results indicate that Cry1 is involved in PhyB-mediated red-light induction of the G-GATA/NOS101 promoter, and that both Cry1 and PhyB are required for effective repression of the GT1/ NOS101 promoter by red or blue light. An interaction between PhyA and PhyB in mediating GT1-GATA/NOS101 promoter light activation was also observed. Furthermore, our data indicate that COP1 and DET1 exert negative control in the dark only on paired LRE promoters but not single LRE promoters. From these results, we conclude that the combinatorial interaction of LREs is essential in determining the ability of light-responsive promoters to be modulated by crucial cellular regulators and to respond to diverse light environments.

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We report here the isolation and characterization of a cotyledon-specific albino locus of Arabidopsis, WHITE COTYLEDONS (WCO). This recessive mutation in the WCO locus, located on the top of Chromosome 1, results in albino cotyledons but green true leaves. An accumulation profile of chlorophylls and ultrastructure of chloroplasts indicate that WCO is necessary for development of functional chloroplasts in cotyledons but is dispensable in true leaves. This was further supported by the fact that the mutants request feeding of sucrose for their survival at the early seedling stage where true leaves have not emerged, but the mutants which have developed true leaves are able to grow autotrophically without sucrose supplementation. The wco mutants accumulate low levels of chloroplast mRNA encoding photosynthesis-related proteins and have a specific defect in 16S rRNA maturation in a cotyledon-specific manner. Although wco mutants exhibited abnormal chloroplasts and chloroplast gene expression in cotyledons, nuclear genes for photosynthetic components are expressed at similar levels to those found in wild-type siblings. This lack of suppression of the nuclear genes is not due to a defect in the signaling of the so-called "plastid factor" to the nucleus since normal suppression of the nuclear genes was observed in response to the photo-oxidative damage due to norflurazon application.

We have identified, purified, and characterized structurally and functionally a 90-kDa immunodominant antigen associated with the water-soluble fraction of Aspergillus fumigatus. This antigen is recognized by 90.3% of serum samples from patients with aspergilloma and should be considered either by itself or better in combination with other purified antigens as a candidate for developing a standardized immunoassay for the detection of aspergilloma. p90 is a glycoprotein containing at least two N-linked sugar chains of 2 and 5 kDa, respectively, which are not necessary for its reactivity with aspergilloma serum samples. Using specific anti-p90 rabbit serum, we have demonstrated that under native conditions, p90 exists in oligomeric form and has associated catalase activity. This activity is resistant to extreme temperatures (>60؇C), reducing agents (40 mM dithiothreitol), high concentrations of denaturing agents such as 8 M urea and 8% sodium dodecyl sulfate, and treatments with ethanol-chloroform-water (5:3:10 [vol/vol]) mixtures.