ABSTRACT
Blue light regulates many physiological and developmental processes in fungi. Most of the blue light responses in the ascomycete Neurospora crassa are dependent on the two blue light regulatory proteins White Collar (WC)-1 and -2. WC-1 has recently been shown to be the first fungal blue light photoreceptor. In the present study, we characterize the Neurospora protein VIVID. VIVID shows a partial sequence similarity with plant blue light photoreceptors. In addition, we found that VIVID non-covalently binds a flavin chromophore. Upon illumination with blue light, VIVID undergoes a photocycle indicative of the formation of a flavin-cysteinyl adduct. VVD is localized in the cytoplasm and is only present after light induction. A loss-of-function vvd mutant was insensitive to increases in light intensities. Furthermore, mutational analysis of the photoactive cysteine indicated that the formation of a flavin-cysteinyl adduct is essential for VIVID functions in vivo. Our results show that VIVID is a second fungal blue light photoreceptor which enables Neurospora to perceive and respond to daily changes in light intensity.
Subject(s)
Fungal Proteins/physiology , Neurospora crassa/physiology , Amino Acid Sequence , Cytoplasm/physiology , Escherichia coli , Fungal Proteins/chemistry , Fungal Proteins/genetics , Light , Molecular Sequence Data , Neurospora crassa/radiation effects , Photoreceptor Cells , Plant Proteins/chemistry , Sequence Alignment , Sequence Homology, Amino Acid , Spectrometry, FluorescenceABSTRACT
The unicellular green alga Haematococcus pluvialis accumulates large amounts of the red ketocarotenoid astaxanthin when exposed to various stress situations such as salt stress and high light intensities. Here, the light regulation of Haematococcus carotenoid biosynthesis was examined. Isolation and characterization of the lycopene beta cyclase gene involved in carotenoid biosynthesis was carried out using a functional complementation approach. Subsequently, gene expression of lycopene cyclase, phytoene synthase, phytoene desaturase and carotenoid hydroxylase was analysed in green flagellate cells. All four genes revealed higher transcript levels in response to increased illumination. Not only the induction of astaxanthin biosynthesis but also carotenoid gene expression was found to be correlated with the redox state of the photosynthetic electron transport. In accordance with this result, increased transcript levels for carotenoid biosynthesis genes were detected under both blue and red light conditions. The application of different inhibitors of the photosynthetic electron flow indicated that the photosynthetic plastoquinone pool functions as the redox sensor for the up-regulation of carotenoid biosynthesis genes. These results suggested that in Haematococcus not only the specific astaxanthin pathway but also general carotenoid biosynthesis is subject to photosynthetic redox control.
Subject(s)
Carotenoids/biosynthesis , Chlorophyta/genetics , beta Carotene/analogs & derivatives , Amino Acid Sequence , Chlorophyta/metabolism , DNA, Complementary/chemistry , DNA, Complementary/genetics , Dose-Response Relationship, Radiation , Gene Expression Regulation/radiation effects , Intramolecular Lyases/genetics , Intramolecular Lyases/metabolism , Light , Molecular Sequence Data , Oxidation-Reduction/radiation effects , Photosynthesis/physiology , Photosynthesis/radiation effects , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Time Factors , Xanthophylls , beta Carotene/biosynthesisABSTRACT
Carotenoids are ubiquitous and essential components of photosynthetic tissues in plants, algae and cyanobacteria. They participate in the light harvesting process and prevent photooxidative damage of the photosynthetic apparatus. Although de-etiolation and growth under different light conditions were reported to have pronounced effects on carotenoid contents in higher plants and algae, very little is known about the light regulation of carotenogenesis on a molecular level. In the present study, we chose the unicellular green alga Chlamydomonas reinhardtii to investigate the regulation of carotenoid biosynthesis genes in response to light. The carotenoid genes phytoene synthase and phytoene desaturase were selected for gene expression studies. Both phytoene synthase and phytoene desaturase revealed a fast up-regulation in response to light, which seemed to be due to transcriptional control. Only blue light was effective whereas illumination with red light did not lead to elevated transcript levels of phytoene synthase and phytoene desaturase. The inhibition of photosynthesis did not abolish the light induction of carotenoid genes. Comparison with published results showed that the carotenoid genes are simultaneously expressed with other genes involved in chlorophyll biosynthesis and light harvesting. This simultaneous expression may represent one mechanism for the coordinated biosynthesis of carotenoids, chlorophylls and the proteins of the photosynthetic apparatus.
