Shedding light on silica biomineralization by comparative analysis of the silica-associated proteomes from three diatom species.

Morphogenesis of the intricate patterns of diatom silica cell walls is a protein-guided process, yet to date only very few such silica biomineralization proteins have been identified. Therefore, it is currently unknown whether all diatoms share conserved proteins of a basal silica forming machinery, and whether unique proteins are responsible for the morphogenesis of species-specific silica patterns. To answer these questions, we extracted proteins from the silica of three diatom species (Thalassiosira pseudonana, Thalassiosira oceanica, and Cyclotella cryptica) by complete demineralization of the cell walls. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis of the extracts identified 92 proteins that we name 'soluble silicome proteins' (SSPs). Surprisingly, no SSPs are common to all three species, and most SSPs showed very low similarity to one another in sequence alignments. In-depth bioinformatics analyses revealed that SSPs could be grouped into distinct classes based on short unconventional sequence motifs whose functions are yet unknown. The results from the in vivo localization of selected SSPs indicates that proteins, which lack sequence homology but share unconventional sequence motifs may exert similar functions in the morphogenesis of the diatom silica cell wall.

ProminTools: shedding light on proteins of unknown function in biomineralization with user friendly tools illustrated using mollusc shell matrix protein sequences.

Biominerals are crucial to the fitness of many organism and studies of the mechanisms of biomineralization are driving research into novel materials. Biomineralization is generally controlled by a matrix of organic molecules including proteins, so proteomic studies of biominerals are important for understanding biomineralization mechanisms. Many such studies identify large numbers of proteins of unknown function, which are often of low sequence complexity and biased in their amino acid composition. A lack of user-friendly tools to find patterns in such sequences and robustly analyse their statistical properties relative to the background proteome means that they are often neglected in follow-up studies. Here we present ProminTools, a user-friendly package for comparison of two sets of protein sequences in terms of their global properties and motif content. Outputs include data tables, graphical summaries in an html file and an R-script as a starting point for data-set specific visualizations. We demonstrate the utility of ProminTools using a previously published shell matrix proteome of the giant limpet Lottia gigantea.

An Efficient Method for the Plating of Haploid and Diploid Emiliania huxleyi on Solid Medium1.

Emiliania huxleyi is a globally important coccolithophore and one of the most successful eukaryotic organisms in the modern oceans. Despite a large body of work on this organism, including the sequencing of its genome, the tools required for forward and reverse functional genetic studies are still undeveloped. Here we present an optimized method for the clonal isolation of E. huxleyi by plating on solid medium. We demonstrate the utility of this method for a variety of strains including haploid, calcifying-diploid, and noncalcifying diploid strains. We show that, in contrast to previous studies, no changes in cell ploidy status occur when the cells are plated. Our method will greatly aid attempts to elucidate the genetic basis of the remarkable physiology of E. huxleyi by forward and reverse genetic approaches.

Exploiting algal mineralization for nanotechnology: bringing coccoliths to the fore.

Complex mineral structures are produced by many microalgal species. Pioneering work on diatom silica has demonstrated the potential of such structures in nanotechnology. The calcified scales of coccolithophores (coccoliths) have received less attention, but the large diversity of architectures make coccoliths attractive as parts for nano-devices. Currently coccolith calcite can be modified by the incorporation of metal ions or adsorption of enzymes to the surface, but genetic modification of coccolithophores may permit the production of coccoliths with customized architectures and surface properties. Further work on the laboratory cultivation of diverse species, the physiochemical properties of coccoliths and on genetic tools for coccolithophores will be necessary to realize the full potential of coccoliths in nanotechnology.

Glucan, Water Dikinase Exerts Little Control over Starch Degradation in Arabidopsis Leaves at Night.

The first step on the pathway of starch degradation in Arabidopsis (Arabidopsis thaliana) leaves at night is the phosphorylation of starch polymers, catalyzed by glucan, water dikinase (GWD). It has been suggested that GWD is important for the control of starch degradation, because its transcript levels undergo strong diel fluctuations, its activity is subject to redox regulation in vitro, and starch degradation is strongly decreased in gwd mutant plants. To test this suggestion, we analyzed changes in GWD protein abundance in relation to starch levels in wild-type plants, in transgenic plants in which GWD transcripts were strongly reduced by induction of RNA interference, and in transgenic plants overexpressing GWD. We found that GWD protein levels do not vary over the diel cycle and that the protein has a half-life of 2 d. Overexpression of GWD does not accelerate starch degradation in leaves, and starch degradation is not inhibited until GWD levels are reduced by 70%. Surprisingly, this degree of reduction also inhibits starch synthesis in the light. To discover the importance of redox regulation, we generated transgenic plants expressing constitutively active GWD. These plants retained normal control of degradation. We conclude that GWD exerts only a low level of control over starch degradation in Arabidopsis leaves.

Interactions between circadian and hormonal signalling in plants.

Growth and development of plants is controlled by external and internal signals. Key internal signals are those generated by hormones and the circadian clock. We highlight interactions between the circadian clock and hormonal signalling networks in regulating the physiology and growth of plants. Microarray analysis has shown that a significant proportion of transcripts involved in hormonal metabolism, catabolism, perception and signalling are also regulated by the circadian clock. In particular, there are interactions between the clock and abscisic acid, auxin, cytokinin and ethylene signalling. We discuss the role of circadian modulation ('gating') of hormonal signals in preventing temporally inappropriate responses. A consideration of the daily changes in physiology provides evidence that circadian gating of hormonal signalling couples the rhythmic regulation of carbon and water utilisation to rhythmic patterns of growth.